CN112831416A

CN112831416A - In-vitro life-sustaining perfusion culture system and control method thereof

Info

Publication number: CN112831416A
Application number: CN202110126464.7A
Authority: CN
Inventors: 王玄; 陈睿
Original assignee: Shanghai Ruiyu Biotech Co Ltd
Current assignee: Shanghai Ruiyu Biotech Co Ltd
Priority date: 2021-01-29
Filing date: 2021-01-29
Publication date: 2021-05-25

Abstract

The invention relates to the technical field of in-vitro life culture, and discloses an in-vitro life maintenance perfusion culture system, which comprises: the premixing unit comprises a premixing cavity, nutrient solution and gas are added into the premixing cavity, and the nutrient solution and the gas are mixed in the premixing cavity to form culture solution; a culture unit comprising a culture chamber, the culture chamber being in communication with the premix chamber; the exchange unit comprises an exchange chamber and a membrane module, the exchange chamber comprises a first exchange cavity and a second exchange cavity, the membrane module is arranged between the first exchange cavity and the second exchange cavity and communicated through the membrane module, the culture chamber and the premixing chamber are both communicated with the first exchange cavity, and the first exchange cavity can receive culture solution flowing out of the culture chamber and can convey the culture solution to the premixing chamber and the culture chamber; and the liquid supplementing unit is communicated with the second exchange cavity and can directionally convey one or more components required by the culture into the second exchange cavity, and the components in the nutrient solution can permeate into the first exchange cavity through the membrane component.

Description

In-vitro life-sustaining perfusion culture system and control method thereof

Technical Field

The invention relates to the technical field of in-vitro life culture, in particular to an in-vitro life maintenance perfusion culture system and a control method thereof.

Background

The organoid is used as a 3D cell tissue, and has quite high accuracy in the aspect of evaluation of the drug effect of an evaluation tool of tumor drugs. At present, the organoid culture method is mainly static culture in a well plate, an operator cuts an obtained primary sample, then coats the primary sample in matrigel at low temperature, then adds a culture medium, puts the culture medium into an incubator for culture, and changes the culture medium 3 times per week and carries out passage once every 7-10 days. This mode of culture presents a number of problems: firstly, the growth speed of organoids is slow, the culture period is long, the culture chamber needs to be opened periodically to change liquid manually in the operation process, and the pollution risk is high; when the number of cultured samples increases, the cost input of manpower, equipment, space and the like is greatly increased. Second, static culture in a culture plate can result in insufficient exchange of organoids with the culture medium, especially the lack of absorption of internal nutrients by the 3D cell tissue, which ultimately limits organoid growth, and limits the extent of organoid growth and the similarity of simulated human organs.

In order to solve the technical problems, the prior art adopts perfusion culture, and culture solution in a culture plate flows out at any time and is discharged, but the perfusion culture mode has low utilization rate of the culture solution, so that the culture cost is high.

Therefore, an in vitro life-support perfusion culture system is needed to solve the above technical problems.

Disclosure of Invention

The invention aims to provide an in vitro life support perfusion culture system and a control method thereof, which can realize the perfusion culture of a culture and the cyclic utilization of a culture solution, improve the utilization rate of the culture solution and reduce the culture cost.

In order to achieve the purpose, the invention adopts the following technical scheme:

in a first aspect, an in vitro life support perfusion culture system is provided, comprising:

the premixing unit comprises a premixing cavity, nutrient solution and gas are added into the premixing cavity, and the nutrient solution and the gas are mixed in the premixing cavity to form culture solution;

the culture unit comprises a culture chamber, the culture chamber is communicated with the premixing chamber, culture solution in the premixing chamber can be conveyed into the culture chamber, and the culture chamber is used for containing a culture and culturing the culture;

the exchange unit comprises an exchange chamber and a membrane assembly, the exchange chamber comprises a first exchange chamber and a second exchange chamber, the membrane assembly is arranged between the first exchange chamber and the second exchange chamber and communicated with each other through the membrane assembly, the first exchange chamber is communicated with the culture chamber, the first exchange chamber can receive culture solution flowing out of the culture chamber and can reflux the culture solution to the culture chamber or the premixing chamber, and the membrane assembly is used for intercepting and/or permeating partial components in the culture solution.

As a preferable technical solution of the in vitro life support perfusion culture system, the pre-mixing unit further includes a fluid infusion chamber, which is communicated with the pre-mixing chamber, and is used for delivering a nutrient fluid into the pre-mixing chamber, and the fluid infusion chamber is capable of directionally delivering one or more components required by the culture into the pre-mixing chamber.

As a preferable technical scheme of the in vitro life support perfusion culture system, a first interface and a second interface are arranged on the first exchange cavity, the first interface is communicated with the culture chamber, and the second interface is communicated with the premixing chamber and/or the culture chamber;

the first exchange cavity can receive the culture solution flowing out of the culture chamber through the first interface, and the first exchange cavity can reflux the culture solution to the culture chamber and/or the premixing chamber through the second interface.

As a preferred technical scheme of the in vitro life support perfusion culture system, the system further comprises a power unit, wherein the power unit is arranged between the premixing chamber and the culture chamber; and/or the presence of a gas in the gas,

the power unit is arranged between the culture chamber and the first interface; and/or the presence of a gas in the gas,

the power unit is arranged between the culture chamber or the premixing chamber and the second interface.

In a second aspect, there is provided an in vitro life support perfusion culture system comprising:

the exchange unit comprises an exchange chamber and a membrane assembly, the exchange chamber comprises a first exchange cavity and a second exchange cavity, the membrane assembly is arranged between the first exchange cavity and the second exchange cavity and is communicated through the membrane assembly, the culture chamber and the premixing chamber are both communicated with the first exchange cavity, the first exchange cavity can receive culture solution flowing out of the culture chamber and can convey the culture solution to the premixing chamber and/or the culture chamber, and the membrane assembly is used for intercepting and/or permeating partial components in the culture solution;

and the liquid supplementing unit is communicated with the second exchange cavity and is used for conveying nutrient solution to the second exchange cavity, the liquid supplementing unit can directionally convey one or more components required by the culture to the second exchange cavity, and at least part of components in the nutrient solution can permeate into the first exchange cavity through the membrane module.

the culture solution in the culture chamber can flow to the first exchange chamber through the first interface, and the culture solution in the first exchange chamber can flow to the premix chamber and/or the culture chamber through the second interface.

As a preferable technical scheme of the in vitro life support perfusion culture system, the system further comprises a power unit, the power unit is arranged between the premixing chamber and the culture chamber, the power unit is arranged between the culture chamber and the first interface, and the power unit is arranged between the premixing chamber and the second interface and/or between the culture chamber and the second interface.

The preferable technical scheme of the in vitro life support perfusion culture system further comprises a collecting unit, wherein the collecting unit is communicated with the second exchange cavity and is used for collecting liquid in the second exchange cavity.

As a preferable technical scheme of the in vitro life support perfusion culture system, at least one of the premixing unit, the culture chamber, the exchange unit and the collection unit is a disposable consumable.

As a preferable technical scheme of the in vitro life support perfusion culture system, the culture chambers are multiple, and the multiple culture chambers are communicated with the premixing chamber;

one of the exchange units is connected to each of the culture chambers, or all of the culture chambers are connected to one of the exchange units.

In a third aspect, an in vitro life support perfusion culture system is provided, comprising:

the temperature control unit comprises a refrigeration assembly and a temperature control module, the refrigeration assembly is electrically connected with the temperature control module, the temperature control module is used for controlling the refrigeration assembly to cool the culture chamber to a first preset temperature, and the first preset temperature is the liquefaction temperature of the culture support structure;

and a sampling port is arranged on the culture chamber and is used for taking out the culture after the culture supporting structure is liquefied.

As a preferable technical scheme of the in vitro life support perfusion culture system, the temperature control unit further includes a heating component, the heating component is electrically connected to the temperature control module, and the temperature control module is configured to control the heating component to heat the culture chamber and/or the premixing chamber.

As an optimal technical scheme of the in vitro life support perfusion culture system, the heating assembly comprises a plurality of heating pieces, the refrigerating assembly comprises a plurality of refrigerating pieces, the heating pieces are arranged at intervals and a plurality of refrigerating pieces are arranged at intervals, one refrigerating piece is arranged between every two adjacent heating pieces, one heating piece is arranged between every two adjacent refrigerating pieces, and the energy of the heating pieces can be transmitted to the refrigerating pieces.

As a preferable technical solution of the in vitro life support perfusion culture system, the temperature control unit further includes a temperature detection element electrically connected to the temperature control module, and the temperature detection element is configured to detect a temperature of the culture solution in the culture chamber.

In a fourth aspect, an in vitro life support perfusion culture system is provided, comprising:

a microscopic observation module comprising an object stage on which the culture chamber is placed and an observation assembly for observing the culture in the culture chamber, the object stage being capable of placing at least one of the culture chambers;

the microscopic observation module further comprises a rack, the objective table is arranged on the rack in a swinging mode, and the objective table can drive the culture chamber to swing synchronously.

As a preferred technical scheme of an in vitro life support perfusion culture system, the microscopic observation module further comprises an autosampler and a sample injection track, the autosampler track is arranged on the frame, the autosampler is slidingly arranged on the sample injection track, and the autosampler is used for injecting samples into the culture chamber.

In a fifth aspect, an in vitro life support perfusion culture system is provided, comprising:

the sterile control module comprises a sterile working chamber, a filtering assembly and a sterilization assembly, at least the culture chamber is arranged in the sterile working chamber, the filtering assembly is used for filtering gas introduced into the sterile working chamber, and the sterilization assembly is used for sterilizing the sterile working chamber.

As a preferred technical scheme of the in vitro life support perfusion culture system, the premixing unit further includes a pH detection element, a dissolved oxygen detection element and a signal detector, the pH detection element and the dissolved oxygen detection element are both disposed in the premixing chamber, and the signal detector can sense signals fed back by the pH detection element and the dissolved oxygen detection element to obtain a pH value and a dissolved oxygen amount of a culture solution in the premixing chamber.

As a preferred technical scheme of the in vitro life support perfusion culture system, the pH detection piece is a pH electrode piece, and the dissolved oxygen detection piece is an dissolved oxygen electrode piece;

the pH electrode plate is arranged on the inner wall of the premixing cavity or in the premixing cavity, and the dissolved oxygen detector is arranged on the inner wall of the premixing cavity or is immersed in liquid in the premixing cavity.

As a preferable technical solution of the in vitro life support perfusion culture system, the premixing unit further includes a gas premixing control unit, which is communicated with the premixing chamber, and is used for delivering gas into the premixing chamber.

As a preferable technical solution of the in vitro life support perfusion culture system, the number of the premixing chambers is plural, the number of the gas premixing control unit is one, and the gas premixing control unit is simultaneously communicated with the plurality of premixing chambers and can control the gas concentration in the plurality of premixing chambers.

As a preferred technical scheme of the in vitro life support perfusion culture system, the culture unit further comprises a mixing module for shaking the culture solution in the culture chamber.

A sixth aspect provides a method for controlling an in vitro life support perfusion culture system, which is directed to the in vitro life support perfusion culture system, and comprises the following steps:

detecting the concentration of the required components of the culture in the culture chamber and/or determining the growth condition of the culture, and controlling the speed of the culture liquid delivered to the culture chamber by the premix chamber according to the detected concentration and/or the determined growth condition of the culture.

In a seventh aspect, a method for controlling an in vitro life support perfusion culture system is provided, which is directed to the in vitro life support perfusion culture system described above, and comprises the following steps:

and detecting the concentration of the components required by the culture in the culture solution of the culture chamber and/or determining the growth condition of the culture, and controlling the speed of the culture solution delivered to the culture chamber by the premix chamber and the speed of the nutrient solution delivered to the second exchange cavity by the solution supplementing unit according to the detected concentration and/or the determined growth condition of the culture.

The invention has the beneficial effects that:

culture solution in the premixing cavity can be conveyed to the culture cavity, the culture solution in the culture cavity can flow out to the first exchange cavity, the culture solution in the first exchange cavity can flow back to the premixing cavity, circulation of the culture solution is achieved, and perfusion culture of a culture is achieved. The membrane module can be with culture medium in the culture solution grow required part of component and be detained in first exchange intracavity, and the waste matter that produces of culture medium metabolism can permeate the membrane module and permeate to the second exchange intracavity in the culture solution, and the culture solution in first exchange intracavity can flow back to the premix chamber, then carries to supply the culture to continue to use in cultivateing the chamber, has also realized the cyclic utilization of culture solution, has improved the utilization ratio of culture solution, has reduced the cultivation cost. And moreover, at least part of components in the nutrient solution can permeate into the first exchange cavity through the membrane module by the nutrient solution provided by the solution supplementing unit, so that the components can be supplemented directionally according to the consumption condition of the components in the culture solution, the use amount of the nutrient solution is reduced, and the culture cost is reduced.

Moreover, the gas premixing control unit can introduce gas into the premixing chamber to realize the adjustment of the dissolved oxygen concentration of the culture solution; the temperature control unit can control the temperature of the premixing cavity and the temperature of the culture cavity, the pH value of the culture solution is adjusted by adding the amount of the nutrient solution and the alkali liquor, and the temperature, the pH value and the dissolved oxygen concentration of the culture solution can be controlled in the premixing cavity, so that the system is free from the constraint of a carbon dioxide incubator, and the culture scale can be enlarged at will.

Drawings

FIG. 1 is a schematic structural diagram of an in vitro life-support perfusion culture system according to an embodiment of the present invention;

fig. 2 is a first schematic structural diagram of a configuration of a temperature control unit according to a first embodiment of the present invention;

fig. 3 is a schematic structural diagram of a configuration of a temperature control unit according to a first embodiment of the present invention;

fig. 4 is a schematic structural diagram of a heating assembly and a cooling assembly according to an embodiment of the present invention;

FIG. 5 is a first schematic structural diagram of a microscopic observation module provided in the first embodiment of the present invention;

FIG. 6 is a schematic structural diagram of a microscopic observation module according to a first embodiment of the present invention;

FIG. 7 is a third schematic structural diagram of a microscopic observation module provided in the first embodiment of the present invention;

FIG. 8 is a schematic structural diagram of a sterility control module according to an embodiment of the present invention;

FIG. 9 is a first schematic structural diagram of a microscopic observation module provided in a third embodiment of the present invention;

FIG. 10 is a schematic structural diagram II of a microscopic observation module provided in a third embodiment of the present invention;

FIG. 11 is a schematic structural diagram of an in vitro life-supporting perfusion culture system provided in the fourth embodiment of the present invention;

FIG. 12 is a schematic structural diagram of an in vitro life-supporting perfusion culture system provided in the fifth embodiment of the present invention;

fig. 13 is a schematic structural diagram of an in vitro life support perfusion culture system provided in the sixth embodiment of the present invention.

In the figure: 1. a pre-mixing unit; 101. a premix chamber; 102. a pH detecting member; 103. a dissolved oxygen detecting member; 104. a fluid infusion chamber; 105. a gas premixing control unit;

2. a culture chamber; 201. a sample addition port; 202. a sampling port; 3. a switching unit; 301. a membrane module; 302. a first exchange chamber; 303. a second exchange chamber;

4. a collection unit; 5. a power unit; 6. a temperature control unit; 61. a heating sheet; 62. a refrigeration plate; 63. a temperature detection member; 7. a blending module;

8. a microscopic observation module; 801. an object stage; 802. a frame; 803. an objective lens; 804. an eyepiece; 805. a first mechanical rail; 806. a second machine rail; 807. a camera; 808. a connecting portion; 809. an autosampler; 810. a sample injection track;

9. a sterility control module; 901. a sterile working chamber; 902. an air inlet fan; 903. a filter assembly; 904. and (5) sterilizing the assembly.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all, embodiments of the present invention. The components of embodiments of the present invention generally described and illustrated in the figures herein may be arranged and designed in a wide variety of different configurations.

Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

It should be noted that: like reference numbers and letters refer to like items in the following figures, and thus, once an item is defined in one figure, it need not be further defined and explained in subsequent figures.

In the description of the present invention, it should be noted that the terms "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings or orientations or positional relationships that are conventionally placed when the products of the present invention are used, and are used only for convenience of describing the present invention and simplifying the description, but do not indicate or imply that the devices or elements to be referred to must have specific orientations, be constructed in specific orientations, and operate, and thus, should not be construed as limiting the present invention. Furthermore, the terms "first," "second," "third," and the like are used solely to distinguish one from another and are not to be construed as indicating or implying relative importance. In the description of the present invention, "a plurality" means two or more unless otherwise specified.

In the description of the present invention, it should also be noted that, unless otherwise explicitly specified or limited, the terms "disposed" and "connected" are to be interpreted broadly, e.g., as being either fixedly connected, detachably connected, or integrally connected; either mechanically or electrically. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.

In the present invention, unless otherwise expressly stated or limited, "above" or "below" a first feature means that the first and second features are in direct contact, or that the first and second features are not in direct contact but are in contact with each other via another feature therebetween. Also, the first feature being "on," "above" and "over" the second feature includes the first feature being directly on and obliquely above the second feature, or merely indicating that the first feature is at a higher level than the second feature. A first feature being "under," "below," and "beneath" a second feature includes the first feature being directly under and obliquely below the second feature, or simply meaning that the first feature is at a lesser elevation than the second feature.

Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the accompanying drawings are illustrative only for the purpose of explaining the present invention, and are not to be construed as limiting the present invention.

Example one

As shown in fig. 1, the present embodiment discloses an in vitro life support perfusion culture system, which includes a premixing unit 1, a culture unit, an exchange unit 3, a collection unit 4, and a power unit 5.

At least one of the premixing unit 1, the culture chamber 2 of the culture unit, the exchange unit 3 and the collection unit 4 is a disposable consumable, and is treated as waste after being used once, so that the sanitary condition in the perfusion culture process can be ensured, and the normal perfusion culture is prevented from being influenced by the pollution of the premixing unit 1, the culture chamber 2, the exchange unit 3 and the collection unit 4.

The premixing unit 1 comprises a premixing chamber 101, a fluid infusion chamber 104 and a gas premixing control unit 105, and nutrient substances required by the growth of cultures such as culture media, factors, medicines and enzymes are contained in nutrient fluid. The fluid infusion chamber 104 is communicated with the premixing chamber 101, nutrient fluid is stored in the fluid infusion chamber 104, and the fluid infusion chamber 104 is used for conveying the nutrient fluid into the premixing chamber 101. A power unit 5 may be disposed between the fluid infusion chamber 104 and the premixing chamber 101 as needed, and the power unit 5 is used to deliver the nutrient fluid in the fluid infusion chamber 104 into the premixing chamber 101 according to the use requirement.

The gas premixing control unit 105 is communicated with the premixing chamber 101, and is used for delivering gas (specifically including oxygen, carbon dioxide, nitrogen and the like) into the premixing chamber 101. Nutrient solution and gas are added into the premixing chamber 101 through the solution supplementing chamber 104 and the gas premixing control unit 105, and the introduced nutrient solution and gas are mixed in the premixing chamber 101 to form culture solution. The oxygen can increase the dissolved oxygen of the culture solution to meet the growth requirement of the culture; the introduced carbon dioxide reduces and adjusts the pH value of the culture solution to meet the growth requirement of the culture. Preferably, in this embodiment, the premixing chamber 101 and the gas premixing control unit 105 are both one, and in other embodiments, a plurality of premixing chambers 101 are provided, and one gas premixing control unit 105 is provided, and the gas premixing control unit 105 is simultaneously communicated with the plurality of premixing chambers 101, so that the gas concentration in the plurality of premixing chambers 101 can be controlled, and specifically, the dissolved oxygen concentration of the culture solution in the premixing chamber 101 can be controlled.

Preferably, the premixing unit 1 further comprises a pH detecting member 102, a dissolved oxygen detecting member 103 and a signal detector, the pH detecting member 102 and the dissolved oxygen detecting member 103 are both disposed in the premixing chamber 101,

the signal detector can sense signals of the pH detector 102 and the dissolved oxygen detector 103, and the sensed signals are calculated and analyzed to obtain the pH value and the dissolved oxygen amount of the culture solution in the premixing chamber 101. With the change of the pH value and the dissolved oxygen amount in the premixing chamber 101, signals detected by the pH detecting element 102 and the dissolved oxygen detecting element 103 may change, specifically, optical signals of the pH detecting element 102 and the dissolved oxygen detecting element 103 may change, and the signal detector may periodically receive the optical signals on the surfaces of the pH detecting element 102 and the dissolved oxygen detecting element 103 to obtain data feedback, and calculate the pH value and the dissolved oxygen amount according to the feedback data. The signal detector can feed detected information back to the liquid supplementing chamber 104 and the gas premixing control unit 105, and if the dissolved oxygen is too high, the liquid supplementing chamber 104 is enabled to convey more nutrient solution into the premixing chamber 101 for dilution so as to reduce the dissolved oxygen of the culture solution in the premixing chamber 101; if the dissolved oxygen is too low, the gas premixing control unit 105 will introduce oxygen into the premixing chamber 101 to increase the dissolved oxygen of the culture solution in the premixing chamber 101. If the pH value is too high, controlling the gas premixing control unit 105 to introduce carbon dioxide so as to reduce the pH value of the culture solution in the premixing chamber 101; if the pH value is too low, the fluid infusion chamber 104 is controlled to deliver more nutrient solution into the premixing chamber 101 for dilution or alkali liquor is introduced into the premixing chamber 101, so as to increase the pH value of the culture solution in the premixing chamber 101.

Specifically, the pH detector 102 is a pH electrode plate, the dissolved oxygen detector 103 is an dissolved oxygen electrode plate, and both are patch materials; the pH electrode plate is arranged on the inner wall of the premixing chamber 101 or in the premixing chamber 101, and the dissolved oxygen detector 103 is arranged on the inner wall of the premixing chamber 101 or in the premixing chamber 101.

The premixing chamber 101 is further provided with an exhaust port, which can exhaust gas in the premixing chamber 101, so as to ensure the stability of the air pressure of the premixing chamber 101. A stirring assembly is provided in the premixing chamber 101, and the stirring assembly can stir the liquid in the premixing chamber 101 to be uniformly mixed and sufficiently immerse the gas into the liquid. An alkali adding port is further formed in the premixing chamber 101, and alkali liquor can be added into the premixing chamber 101 through the alkali adding port to adjust the pH value of the culture solution in the premixing chamber 101.

The nutrient solution is premixed with oxygen and the like in the premixing chamber 101 to form a culture solution, and the culture solution formed by mixing the nutrient solution with the oxygen and the like is more suitable for the growth of the culture by controlling corresponding variables (such as pH value and dissolved oxygen), so that nutrients required for the growth can be sufficiently provided for the culture. Compared with the prior art in which nutrient solution, oxygen and the like are directly introduced into the culture chamber, the design of complex pipelines, valves and detection pieces is saved, and the system cost is reduced.

The culture unit comprises a culture chamber 2, the culture chamber 2 is communicated with a premixing chamber 101, and culture solution in the premixing chamber 101 can be conveyed into the culture chamber 2. Whether to arrange power unit 5 between premix chamber 101 and culture chamber 2 may be determined according to the relative arrangement positions of premix chamber 101 and culture chamber 2 and the requirements of the transfer speed and transfer amount of the culture solution, and culture solution in premix chamber 101 is transferred into culture chamber 2 using power unit 5. Preferably, in the present embodiment, the culture chambers 2 are one, that is, one premix chamber 101 corresponds to one culture chamber. In other embodiments, there are a plurality of culture chambers 2, and a plurality of culture chambers 2 are all communicated with premix chamber 101, that is, a plurality of culture chambers 2 are connected in parallel, either one exchange unit 3 may be connected to each culture chamber 2, all culture chambers 2 may be connected to one exchange unit, or several culture chambers 2 may supply one exchange unit 3. The device can be used for simultaneously culturing a plurality of cultures, so that the consistency of the cultures in the subsequent test process can be conveniently ensured, and the accuracy of test result comparison can be ensured.

The culture chamber 2 is used for containing a culture and culturing the culture. Specifically, the culture chamber 2 is provided with a sample addition port 201 and a sampling port 202. The culture can be added to the culture chamber 2 through the sample addition port 201. In addition, growth factors, drugs and the like can be added into the culture chamber 2 through the sample adding port 201 to promote or inhibit the growth of the culture according to experimental requirements, and enzymes, matrigel treatment reagents, staining reagents and the like can be added into a system after the culture is completed for collecting, treating or characterizing the culture. Samples of the culture or broth can be taken for analysis during the incubation through the sampling port 202.

The exchange unit 3 comprises an exchange chamber and a membrane module 301, the exchange chamber comprises a first exchange chamber 302 and a second exchange chamber 303, the membrane module 301 is arranged between the first exchange chamber 302 and the second exchange chamber 303, and is communicated through the membrane module 301, the first exchange chamber 302 can receive the culture solution flowing out of the culture chamber 2 and can reflux the culture solution to the culture chamber 2, and the membrane module 301 is used for intercepting and/or permeating part of components in the culture solution. Specifically, the membrane module 301 can trap nutrient components required by the culture in the culture solution in the first exchange cavity 302, such as serum, growth factors, enzymes and the like, and metabolites generated by the growth of the culture in the first exchange cavity 302, such as urea, carbon dioxide and the like, can permeate through the membrane module 301 into the second exchange cavity 303 and then be discharged from the second exchange cavity 303.

The exchange unit 3 and the membrane module 301 are arranged, so that useful components in the culture solution can be intercepted while perfusion culture is realized, and the useful components flow back to the culture chamber 2 for absorption and utilization again, the utilization rate of the culture solution is improved, and the culture cost is reduced. The exchange unit 3 is arranged outside the culture chamber 2, so that the effective area of the membrane module 301 can be set to be larger and the structure is more flexible, and the exchange efficiency is further improved.

The collecting unit 4 is communicated with the second exchange cavity 303 and is used for collecting the culture solution in the second exchange cavity 303. Whether the power unit 5 is arranged between the second exchange cavity 303 and the collection unit 4 or not can be determined according to the relative arrangement positions of the second exchange cavity 303 and the collection unit 4 and the requirements of the culture liquid conveying speed and conveying amount, and preferably, in the embodiment, the power unit 5 is arranged between the collection unit 4 and the second exchange cavity 303, and the power unit 5 is used for conveying the culture liquid in the second exchange cavity 303 to the collection unit 4.

Specifically, the first exchange cavity 302 is provided with a first interface and a second interface, and both the first interface and the second interface are communicated with the culture chamber 2. The first exchange chamber 302 can receive the culture solution flowing out of the culture chamber 2 through the first interface, and the first exchange chamber 302 can reflux the culture solution to the culture chamber 2 through the second interface. Preferably, in the present embodiment, the power unit 5 is provided between the culture chamber 2 and the first port and between the culture chamber 2 and the second port. The power unit 5 between the culture chamber 2 and the first interface can convey the culture solution in the culture chamber 2 to the first exchange cavity 302 through the first interface; the power unit 5 between the culture chamber 2 and the second interface can transfer the culture solution in the first exchange cavity 302 to the culture chamber 2 through the second interface.

Because the consumption speed of each component in the culture solution is different in the culture process, one or more components can be added directionally through the solution supplementing chamber 104 in the scheme of the embodiment, the balance of each component is ensured, the whole solution changing step is reduced, the cost is reduced, and the culture pollution probability is reduced. The liquid supplementing chamber 104 supplements liquid to the premixing chamber 101, and then the premixing chamber 101 delivers nutrient solution to the culture chamber 2; therefore, the concentration and composition of the nutrient solution in the fluid replacement chamber 104 do not have to be adjusted to be the same as the concentration and composition of the culture solution required for the culture, and the volume of the fluid replacement chamber 104 can be made smaller. Meanwhile, the perfusion rate of the premix chamber 101 may be higher without worrying about the excessively fast consumption of the culture solution due to the increase of the perfusion rate, improving the culture efficiency, increasing the variety of the cultures that can be cultured.

Specifically, the concentration of the culture liquid component flowing out of the culture chamber 2 can be detected and supplemented with the replenishment chamber 104 in a targeted manner. The concentration detection can adopt sampling detection or in-situ detection (such as infrared spectrum technology and fluorescence detection technology). Preferably, the present embodiment employs fluorescence detection techniques. The system can also be provided with an automatic sample adding module for automatically adding nutrient components into the fluid infusion chamber 104 based on the concentration detection result.

Preferably, the culture unit further comprises a temperature control unit 6, wherein the temperature control unit 6 is used for controlling the temperature of the premixing chamber 101 and the culture chamber 2, and specifically, the premixing chamber 101 and the culture chamber 2 can be heated to raise the temperature of the culture solution in the premixing chamber 101 and the temperature of the culture solution in the culture chamber 2 to a physiological temperature suitable for the growth of the culture, so as to culture the culture; when the temperature is too high, the temperature of the premixing chamber 101 and the culture chamber 2 may be cooled and lowered to lower the temperature of the culture solution in the culture chamber 2 of the premixing chamber 101 to a physiological temperature suitable for the growth of the culture, so as to perform the culture of the culture. After the culture is finished, when matrigel needs to be removed and the culture needs to be collected, the temperature control unit 6 can control the temperature of the culture solution in the culture chamber 2 to a first preset temperature, wherein the first preset temperature is the liquefaction temperature of the culture support structure, and the culture can be taken out through the sampling port 202 after the culture support structure is liquefied. The culture support structure forms a network support for the culture that facilitates 3D growth of the culture. Culture support structures include, but are not limited to, matrigel (2-8 degrees liquefied), synthetic gel (room temperature 25 degrees liquefied).

Specifically, temperature control unit 6 includes heating assembly, refrigeration subassembly and temperature control module, and heating assembly and refrigeration subassembly all are connected with temperature control module electricity, and temperature control module is used for controlling heating assembly heating and cultivates cavity 2, still is used for controlling refrigeration subassembly cooling and cultivates cavity 2.

The outer walls of the premixing chamber 101 and the culture chamber 2 are provided with a heating assembly and a refrigerating assembly, the heating assembly comprises a plurality of heating sheets 61, and the heating sheets 61 are arranged on the outer wall of the culture chamber 2. The heating sheet 61 may be a transparent heating plate, a metal heating plate, a semiconductor heating plate, a heating blanket, a heating sheet, or the like. The refrigeration assembly comprises a plurality of refrigeration pieces 62, the refrigeration pieces 62 are arranged on the outer wall of the culture chamber 2, and the refrigeration pieces 62 can be semiconductor refrigeration pieces 62 or other refrigeration materials.

The heating and

cooling plates

61, 62 may be located at different positions of the incubation chamber 2, such as: as shown in fig. 2, the heating sheet 61 is positioned at the bottom of the culture chamber 2, and the cooling sheet 62 is positioned at the side of the culture chamber 2; alternatively, as shown in FIG. 3, the heating sheet 61 is located at the side of the incubation chamber 2, and the cooling sheet 62 is located at the bottom of the incubation chamber 2. Of course, the heating plate 61 and the cooling plate 62 may also be located at the same position of the cultivation chamber 2, for example, both located at the same side or bottom of the cultivation chamber 2.

Preferably, in this embodiment, as shown in fig. 4, the heating sheet 61 and the cooling sheet 62 are located on the same side or bottom wall of the incubation chamber 2. Specifically, a plurality of heating sheets 61 are arranged at intervals, a plurality of cooling sheets 62 are arranged at intervals, one cooling sheet 62 is arranged between two adjacent heating sheets 61, one heating sheet 61 is arranged between two adjacent cooling sheets 62, the heating sheets 61 are in close contact with the cooling sheets 62 adjacent to the heating sheets 61, the cooling sheets 62 are in close contact with the heating sheets 61 adjacent to the cooling sheets 62, and energy of the heating sheets 61 can be transmitted to the cooling sheets 62. The heating plate 61 and the refrigerating plate 62 are made of materials with good heat conduction performance, and can be structurally processed into an inlaid structure, so that the heating plate 61 can be embedded between the refrigerating plates 62, and the refrigerating plates 62 can be embedded between the heating plates 61.

When needing to cultivate cavity 2 heating and rising temperature, heat piece 61 and heat to appointed temperature under temperature control module's control, because refrigeration piece 62 is the heat conduction material, and the heat of heating piece 61 can transmit to refrigeration piece 62, heats refrigeration piece 62 when heating to cultivateing cavity 2, the temperature of refrigeration piece 62 rises to appointed temperature rapidly, then cultivates cavity 2 by heating piece 61 and refrigeration piece 62 common heating. When needing to cultivate cavity 2 cooling, refrigeration piece 62 is cooled down to the assigned temperature under temperature control module's control, because heat piece 61 is the heat conduction material, and heat piece 61's heat can transmit to refrigeration piece 62, and refrigeration piece 62 is to heating piece 61 cooling when cultivateing cavity 2 cooling, makes its temperature drop to the assigned temperature rapidly, then cultivates cavity 2 by refrigeration piece 62 and the common cooling of heating piece 61.

The heating sheets 61 and the refrigerating sheets 62 are arranged in a staggered mode and can transfer heat, the heating sheets and the refrigerating sheets can heat and cool the culture chamber 2 together, the heat exchange area is increased, and the heat exchange efficiency is improved.

The temperature control unit 6 further comprises a temperature detection member 63, which may be a temperature sensor. The temperature detection piece 63 is electrically connected with the temperature control module, the temperature detection piece 63 can be arranged on the inner wall of the culture cavity 2 or on the outer wall of the culture cavity 2 and used for detecting the temperature of the culture solution in the culture cavity and feeding back the detection value to the temperature control module, and the temperature control module compares the temperature value detected by the temperature detection piece 63 with an actually required set value so as to control the heating assembly to heat the temperature culture cavity 2 or control the refrigerating assembly to cool the culture cavity 2.

The heating and cooling modules and the temperature detector 63 of the premix chamber 101 are constructed and arranged in the same manner as those of the incubation chamber 2, and will not be described in detail.

Optionally, with continued reference to fig. 1, the culture unit further comprises a mixing module 7, the mixing module 7 being embodied as a mechanical shaking mechanism for shaking the culture fluid in the culture chamber 2, in particular by shaking the culture chamber 2, to shake the culture fluid in the culture chamber 2, providing a more sufficient exchange opportunity for the culture fluid and the culture. The shaking amplitude and speed of the mixing module 7 can be matched with the speed of the culture solution delivered into the culture chamber 2, for example, a higher culture solution delivery speed can promote the exchange opportunity of the culture solution and the culture but can cause the culture solution to be excessively consumed. The lower culture solution conveying speed is matched with the large mechanical shaking, so that the aims of updating the culture solution, increasing the exchange chance of the nutrient solution and the culture and promoting the exchange of nutrient substances can be fulfilled simultaneously.

Optionally, as shown in fig. 5 and 6, the culture unit further comprises a microscopic observation module 8 comprising an object stage 801 and an observation assembly, the culture chamber 2 being placed on the object stage 801, in particular the culture chamber 2 being placed on the homogenisation module 7, the homogenisation module 7 being placed on the object stage 801. The object stage 801 can be used for placing at least one culture chamber 2, in the embodiment, a placing station is arranged on the object stage 801, and the blending module 7 is placed on the placing station; in other embodiments, the stage 801 is provided with a plurality of placement stations (see fig. 7 in particular), and each placement station can be provided with one blending module 7.

The observation assembly is for observing the culture within the culture chamber 2, and specifically includes an objective lens 803, an eyepiece lens 804, and a camera 807 (specifically a CCD or CMOS camera). The microscopic observation module 8 further includes a frame 802, an objective lens 803 is provided on the frame 802 so as to be movable in the vertical direction, and an eyepiece lens 804 is also provided on the frame 802 so that the culture in the culture chamber 2 can be observed through the eyepiece lens 804.

The frame 802 is provided with a first mechanical rail 805, and a second mechanical rail 806 is slidably provided on the first mechanical rail 805, wherein the first mechanical rail 805 and the second mechanical rail 806 are perpendicular to each other and are both located in a horizontal plane. The stage 801 is slidably disposed on the second mechanical rail 806, and the stage 801 can be adjusted to any position in the horizontal plane by sliding the stage 801 relative to the second mechanical rail 806 and sliding the second mechanical rail 806 relative to the first mechanical rail 805 so that the objective lens 803 faces the observation chamber, and the culture in the culture chamber 2 is focused using the eyepiece 804 to observe the culture. If image data is to be recorded during observation, the image data may be photographed using the camera 807.

Preferably, as shown in fig. 7, the microscopic observation module 8 further includes an autosampler 809 and a sample rail 810, wherein the autosampler rail 810 is disposed on the rack 802, the autosampler 809 is slidably disposed on the sample rail 810, and the autosampler 809 is driven by the sample driving member to slide on the sample rail 810 for sample feeding. The autosampler 809 is mainly used for adding the culture, the reagent or the medicine in the culture chamber 2, and the autosampler 809 is used for adding the sample, so that the manual sample adding is avoided, the sanitary condition of the culture chamber 2 is ensured, and the culture is not easy to be polluted.

Optionally, as shown in fig. 8, the in vitro life support perfusion culture system further includes a sterile control module 9, which includes a sterile working chamber 901, a filter assembly 903 and a sterilization assembly 904, at least the culture chamber 2 is disposed in the sterile working chamber 901, in this embodiment, the premixing unit 1, the culture unit, the exchange unit 3, the collection unit 4, the power unit 5, and the like are disposed in the sterile working chamber 901, that is, all the parts of the culture system of the culture except the sterile control module 9 are located in the sterile working chamber 901.

The sterile working room 901 is provided with a vent, the filter element 903 is arranged at the vent, the vent is provided with an air inlet fan 902, air is introduced into the sterile working room 901 through the air inlet fan 902, and the filter element 903 is used for filtering the air introduced into the sterile working room 901, so that dust, magazines and the like cannot enter the sterile working room 901. The sterilization assembly 904 is used for sterilizing and disinfecting the sterile working chamber 901, specifically, the sterilization assembly 904 is an ultraviolet lamp and is arranged at an air inlet, the sterilization assembly 904 can sterilize the sterile working chamber 901, and can also sterilize air introduced into the sterile working chamber 901, so that the sterile air can be prevented from entering the sterile working chamber 901.

The embodiment also discloses a control method of the in vitro life support perfusion culture system, which aims at the in vitro life support perfusion culture system and comprises the following steps:

the concentration of each component required by the culture in the culture solution in the culture chamber 2 is detected and/or the growth condition of the culture is determined, the speed of the premix chamber 101 for delivering the culture solution to the culture chamber 2 is controlled according to the detected concentration and/or the determined growth condition of the culture, and meanwhile, the speed of the solution supplementing chamber 104 for delivering the nutrient solution to the premix chamber 1 is controlled, and the speed of the gas premix control unit 105 for delivering gas to the premix chamber 101 is controlled.

Example two

The embodiment discloses an in vitro life support perfusion culture system, which comprises a premixing unit 1, a culture unit, an exchange unit 3, a collection unit 4 and a power unit 5.

The in vitro life support perfusion culture system in this embodiment has a structure substantially the same as that of the in vitro life support perfusion culture system in the first embodiment, except that the culture unit does not include the temperature control unit 6 and the blending unit, and the culture unit in this embodiment further includes a blending temperature control module for controlling the temperature of the culture chamber 2 and shaking the culture solution in the culture chamber 2.

The blending temperature control module comprises a mechanical shaking mechanism and a temperature control assembly, and the mechanical shaking mechanism is provided with a mounting surface. The temperature control assembly is used for controlling the temperature of the culture chamber 2, and specifically can heat the premixing chamber 101 and the culture chamber 2 to enable the temperature of the culture solution in the premixing chamber 101 and the temperature of the culture solution in the culture chamber 2 to be raised to a physiological temperature suitable for the growth of the culture so as to culture the culture; after the culture is finished and the matrigel is required to be removed and the culture is collected, the temperature control assembly can make the culture solution in the culture chamber 2 drop to the matrigel liquefaction temperature in the culture solution so as to collect the culture. Specifically, accuse temperature subassembly includes heating subassembly, refrigeration subassembly and temperature control module, and heating subassembly and refrigeration subassembly all are connected with temperature control module electricity, and temperature control module is used for controlling heating subassembly heating and cultivates cavity 2, still is used for controlling refrigeration subassembly cooling and cultivates cavity 2.

The heating member includes a plurality of heating sheets 61, and the heating sheets 61 are provided on the outer wall of the incubation chamber 2. The heating sheet 61 may be a transparent heating plate, a metal heating plate, a semiconductor heating plate, a heating blanket, a heating sheet, or the like. The refrigeration assembly comprises a plurality of refrigeration pieces 62, the refrigeration pieces 62 are arranged on the outer wall of the culture chamber 2, and the refrigeration pieces 62 can be semiconductor refrigeration pieces 62 or other refrigeration materials. Specifically, a plurality of heating sheets 61 are arranged at intervals, a plurality of cooling sheets 62 are arranged at intervals, one cooling sheet 62 is arranged between two adjacent heating sheets 61, one heating sheet 61 is arranged between two adjacent cooling sheets 62, the heating sheets 61 are in close contact with the cooling sheets 62 adjacent to the heating sheets 61, the cooling sheets 62 are in close contact with the heating sheets 61 adjacent to the cooling sheets 62, and energy of the heating sheets 61 can be transmitted to the cooling sheets 62. The heating plate 61 and the refrigerating plate 62 are made of materials with good heat conduction performance, and can be structurally processed into an inlaid structure, so that the heating plate 61 can be embedded between the refrigerating plates 62, and the refrigerating plates 62 can be embedded between the heating plates 61. The heating sheets 61 and the refrigerating sheets 62 are sequentially arranged on the installation surface and are mainly used for heating or cooling the bottom surface of the culture chamber 2, and further controlling the temperature of the culture solution in the culture chamber 2.

The temperature control assembly further comprises a temperature detection member 63, which may be a temperature sensor. The temperature detection piece 63 is electrically connected with the temperature control module, the temperature detection piece 63 can be arranged on the inner wall of the culture cavity 2 or on the outer wall of the culture cavity 2 and used for detecting the temperature of the culture solution in the culture cavity and feeding back the detection value to the temperature control module, and the temperature control module compares the temperature value detected by the temperature detection piece 63 with an actually required set value so as to control the heating assembly to heat the temperature culture cavity 2 or control the refrigerating assembly to cool the culture cavity 2.

EXAMPLE III

The in vitro life support perfusion culture system in this embodiment has basically the same structure as the in vitro life support perfusion culture system in the first embodiment, except that the culture unit in this embodiment does not include the blending module 7, and the microscopic observation unit has a different structure. As shown in fig. 9, the microscopic observation module 8 in the present embodiment includes a stage 801, an observation unit, and a frame 802, the culture chamber 2 is placed on the stage 801, the stage 801 is swingably provided on the frame 802, and the swing stage 801 is capable of shaking the culture solution in the culture chamber 2.

The culture chamber 2 is placed on the stage 801, and the culture chamber 2 is placed on the stage 801. The object stage 801 can be used for placing at least one culture chamber 2, in this embodiment, a placing station is arranged on the object stage 801, and the culture chamber 2 is placed on the placing station; in other embodiments, the stage 801 is provided with a plurality of placing stations, each of which can place one culture chamber 2.

The observation assembly is for observing the culture in the culture chamber 2, and specifically, the observation assembly includes an objective lens 803, an eyepiece lens 804, and a camera 807 (specifically, a CCD camera). The microscopic observation module 8 further includes a frame 802, an objective lens 803 is provided on the frame 802 so as to be movable in the vertical direction, and an eyepiece lens 804 is also provided on the frame 802 so that the culture in the culture chamber 2 can be observed through the eyepiece lens 804.

The frame 802 is provided with a first mechanical rail 805, and a second mechanical rail 806 is slidably provided on the first mechanical rail 805, wherein the first mechanical rail 805 and the second mechanical rail 806 are perpendicular to each other and are both located in a horizontal plane. A connecting portion 808 is attached to the bottom of the stage 801, a rotating shaft is provided on the connecting portion 808, the stage 801 is rotatably connected to the rotating shaft, the stage 801 can swing with respect to the connecting portion 808, and the connecting portion 808 is slidably provided on the second machine rail 806 and can slide in the longitudinal direction of the second machine rail 806. The microscopic observation module 8 further comprises a driving member, an output end of which is connected to the object stage 801, for driving the object stage 801 to shake the culture solution in the culture chamber 2. Provides more sufficient exchange opportunity for the culture solution and the culture. The amplitude and speed of the shaking of the stage 801 can be matched to the speed of the culture medium being fed into the culture chamber 2, for example, a higher rate of the feeding of the culture medium can promote the chance of exchanging the culture medium with the culture medium, but can result in excessive consumption of the culture medium. The lower culture solution conveying speed is matched with the large mechanical shaking, so that the aims of updating the culture solution, increasing the exchange chance of the nutrient solution and the culture and promoting the exchange of nutrient substances can be fulfilled simultaneously.

The adjustment of the stage 801 to any position in the horizontal plane can be achieved by sliding the stage 801 relative to the second mechanical rail 806 and sliding the second mechanical rail 806 relative to the first mechanical rail 805 so that the objective lens 803 is directed to the observation room and the culture in the culture chamber 2 is focused using the eyepiece 804 to observe the culture. If image data is to be recorded during observation, the image data may be photographed using the camera 807. The stage 801 may be shaken during the observation to bring the culture into a better observation state for easy observation.

As shown in fig. 10, the microscopic observation module 8 preferably further includes an autosampler 809 and a sample rail 810, wherein the autosampler rail 810 is disposed on the rack 802, the autosampler 809 is slidably disposed on the sample rail 810, and the autosampler 809 is driven by the sample driving member to slide on the sample rail 810 for sample feeding. The autosampler 809 is mainly used for adding the culture, the reagent or the medicine in the culture chamber 2, and the autosampler 809 is used for adding the sample, so that the manual sample adding is avoided, the sanitary condition of the culture chamber 2 is ensured, and the culture is not easy to be polluted.

Example four

As shown in fig. 11, the present embodiment discloses an in vitro life support perfusion culture system, which includes a premixing unit 1, a culture unit, an exchange unit 3, a collection unit 4, and a power unit 5.

The in vitro life support perfusion culture system in this embodiment has substantially the same structure as the in vitro life support perfusion culture system in the first embodiment, except that the first exchange chamber 302 directly supplies the culture solution to the premix chamber 101 without directly supplying the culture solution to the culture chamber 2, and the power unit 5 is provided between the first exchange chamber 302 and the premix chamber 101, and the culture solution in the second exchange chamber 302 is transferred to the premix chamber 101 by using the power unit 5.

EXAMPLE five

As shown in fig. 12, the present embodiment discloses an in vitro life support perfusion culture system, which includes a premixing unit 1, a culture unit, an exchange unit 3, a fluid infusion unit 11, a collection unit 4, and a power unit 5.

The premixing unit 1 includes a premixing chamber 101 and a gas premixing control unit 105, and the culture solution contains nutrients necessary for the growth of the culture such as a culture medium, factors, drugs, and enzymes. The gas premixing control unit 105 is communicated with the premixing chamber 101, and is used for delivering gas (specifically including oxygen, carbon dioxide, nitrogen and the like) into the premixing chamber 101. The premix chamber 101 may receive a nutrient solution, and the manner of receiving the nutrient solution will be described later and will not be described in detail herein. Gas is added into the premixing chamber 101 through the gas premixing control unit 105, and the introduced nutrient solution and the gas are mixed in the premixing chamber 101 to form a culture solution. The oxygen can increase the dissolved oxygen of the culture solution to meet the growth requirement of the culture; the introduced carbon dioxide reduces and adjusts the pH value of the culture solution to meet the growth requirement of the culture. Preferably, in this embodiment, the premixing chamber 101 and the gas premixing control unit 105 are both one, and in other embodiments, a plurality of premixing chambers 101 are provided, and one gas premixing control unit 105 is provided, and the gas premixing control unit 105 is simultaneously communicated with the plurality of premixing chambers 101, so that the gas concentration in the plurality of premixing chambers 101 can be controlled, and specifically, the dissolved oxygen concentration of the culture solution in the premixing chamber 101 can be controlled.

Preferably, the signal detector can sense signals of the pH detector 102 and the dissolved oxygen detector 103, and the sensed signals are calculated and analyzed to obtain the pH value and the dissolved oxygen amount of the culture solution in the premixing chamber 101. With the change of the pH value and the dissolved oxygen amount in the premixing chamber 101, signals detected by the pH detecting element 102 and the dissolved oxygen detecting element 103 may change, specifically, optical signals of the pH detecting element 102 and the dissolved oxygen detecting element 103 may change, and the signal detector may periodically receive the optical signals on the surfaces of the pH detecting element 102 and the dissolved oxygen detecting element 103 to obtain data feedback, and calculate the pH value and the dissolved oxygen amount according to the feedback data. The signal detector can feed back the detected information to the gas premixing control unit 105, and if the dissolved oxygen is too low, the gas premixing control unit 105 introduces oxygen into the premixing chamber 101 to increase the dissolved oxygen of the culture solution in the premixing chamber 101. If the pH value is too high, the gas premixing control unit 105 is controlled to introduce carbon dioxide to reduce the pH value of the culture solution in the premixing chamber 101. And if the pH value is too low, introducing alkali liquor into the premixing chamber 101 to increase the pH value of the culture solution in the premixing chamber 101.

The culture unit comprises a culture chamber 2, the culture chamber 2 is communicated with a premixing chamber 101, and culture solution in the premixing chamber 101 can be conveyed into the culture chamber 2. Whether the power unit 5 is arranged between the premixing chamber 101 and the culture chamber 2 or not can be determined according to the relative arrangement positions of the premixing chamber 101 and the culture chamber 2 and the requirements of the culture liquid conveying speed and the conveying amount, and the culture liquid in the premixing chamber 101 is conveyed into the culture chamber 2 by using the power unit 5. Preferably, in the present embodiment, the culture chambers 2 are one, that is, one premix chamber 101 corresponds to one culture chamber. In other embodiments, there are a plurality of culture chambers 2, and a plurality of culture chambers 2 are all communicated with premix chamber 101, that is, a plurality of culture chambers 2 are connected in parallel, either one exchange unit 3 may be connected to each culture chamber 2, all culture chambers 2 may be connected to one exchange unit, or several culture chambers 2 may supply one exchange unit 3. The device can be used for simultaneously culturing a plurality of cultures, so that the consistency of the cultures in the subsequent test process can be conveniently ensured, and the accuracy of test result comparison can be ensured.

The exchange unit 3 comprises an exchange chamber and a membrane module 301, the exchange chamber comprises a first exchange cavity 302 and a second exchange cavity 303, the membrane module 301 is arranged between the first exchange cavity 302 and the second exchange cavity 303 and is communicated through the membrane module 301, and the membrane module 301 is used for intercepting and/or permeating partial components in the culture solution. Specifically, the membrane module 301 can trap useful components in the culture solution in the first exchange cavity 302, such as serum, growth factors, enzymes, and the like, in the first exchange cavity 302, and metabolites, such as urea, carbon dioxide, and the like, generated by the growth of the culture in the first exchange cavity 302 can permeate through the membrane module 301 into the second exchange cavity 303 and then be discharged from the second exchange cavity 303.

Cultivate cavity 2 and premix chamber 101 and all be linked together with first swap cavity 302, first swap cavity 302 can enough receive the culture solution that cultivates cavity 2 and flow out, can be to the palirrhea culture solution of premix chamber 101 again, specifically, is provided with first interface and second interface on the first swap cavity 302, first interface with cultivate cavity 2 intercommunication, the second interface is linked together with premix chamber 101. The first swap chamber 302 can receive the culture solution flowing out of the culture chamber 2 through the first interface, and the first swap chamber 302 can reflux the culture solution to the premix chamber 101 through the second interface. Preferably, in the present embodiment, the power unit 5 is provided between the culture chamber 2 and the first port and between the premix chamber 101 and the second port. The power unit 5 between the culture chamber 2 and the first interface can convey the culture solution in the culture chamber 2 to the first exchange cavity 302 through the first interface; the power unit 5 between the premixing chamber 101 and the second interface can transfer the culture solution in the first exchange cavity 302 to the premixing chamber 101 through the second interface.

The exchange unit 3 and the membrane module 301 are arranged, so that useful components in the culture solution can be intercepted while perfusion culture is realized, the useful components flow back to the premixing chamber 101 and are conveyed to the culture chamber 2 for absorption and utilization again, the utilization rate of the culture solution is improved, and the culture cost is reduced. The exchange unit 3 is arranged outside the culture chamber 2, so that the effective area of the membrane module 301 can be set to be larger and the structure is more flexible, and the exchange efficiency is further improved.

The collecting unit 4 is communicated with the second exchange cavity 303 and is used for collecting the culture solution in the second exchange cavity 303. Whether the power unit 5 is arranged between the second exchange cavity 303 and the collection unit 4 or not can be determined according to the relative arrangement positions of the second exchange cavity 303 and the collection unit 4 and the requirements of the culture liquid conveying speed and conveying amount, and preferably, in the embodiment, the power unit 5 is arranged between the collection unit 4 and the second exchange cavity 303, and the culture liquid in the second exchange cavity 303 is conveyed to the collection unit 4 by using the power unit 5.

The liquid supplementing unit 11 is communicated with the second exchange cavity 303 and is used for delivering nutrient solution to the second exchange cavity 303, and at least part of components in the nutrient solution in the second exchange cavity 303 can permeate into the first exchange cavity 302 through the membrane component 301. Whether the power unit 5 is arranged between the liquid supplementing unit 11 and the second exchange cavity 303 can be determined according to the relative arrangement positions of the second exchange cavity 303 and the liquid supplementing unit 11 and the requirements of the conveying speed and the conveying amount of the nutrient solution, and preferably, the power unit 5 is arranged between the liquid supplementing unit 11 and the second exchange cavity 303 in the embodiment, and the nutrient solution is conveyed into the second exchange cavity 303 by using the power unit 5. The concentration of the nutrient solution is greater than that of the culture solution in the first exchange cavity 302, and the concentration difference between the two can drive the components in the nutrient solution to move into the first exchange cavity 302 through the membrane module 301, and then the components are conveyed to the nutrient solution in the premixing chamber 101, and the nutrient solution and the gas are mixed to form the culture solution in the premixing chamber 101.

Because the consumption speed of each component in the culture solution is different in the culture process, one or more components can be added directionally through the solution supplementing unit 11 in the scheme of the embodiment, the balance of each component is guaranteed, the whole solution changing step is reduced, the cost is reduced, and the culture pollution probability is reduced. The liquid supplementing unit 11 supplements liquid to the second exchange cavity 303, and then conveys nutrient solution to the culture cavity 2 from the first exchange cavity 302; therefore, the concentration and composition of the nutrient solution in the solution supplementing unit 11 do not have to be adjusted to be the same as those of the culture solution required for the culture, and the volume of the solution supplementing unit 11 can be made smaller. Meanwhile, the perfusion rate of the premix chamber 101 may be higher without worrying about the excessively fast consumption of the culture solution due to the increase of the perfusion rate, improving the culture efficiency, increasing the variety of the cultures that can be cultured.

Specifically, the concentration of the culture liquid component flowing out of the culture chamber 2 can be detected and supplemented with the solution supplementing unit 11 in a targeted manner. The concentration detection can adopt sampling detection or in-situ detection (such as infrared spectrum technology and fluorescence detection technology). Preferably, the present embodiment employs fluorescence detection techniques. The system can also be provided with an automatic sample adding module, and nutrient components are automatically added into the liquid supplementing unit 11 based on the concentration detection result.

Preferably, the culture unit further comprises a temperature control unit 6 and a microscopic observation module 8, and the temperature control unit 6 and the microscopic observation module 8 have the same structure as the temperature control unit 6 and the microscopic observation module 8 in the first embodiment, and are not described in detail herein.

Optionally, the in vitro life support perfusion culture system further comprises a sterile control module 9, and the sterile control module 9 has the same structure as the sterile control module 9 in the first embodiment, and will not be described in detail herein.

the concentration of the components required by the culture in the culture solution in the culture chamber 2 is detected and/or the growth condition of the culture is determined, and the speed of the premix chamber 101 for delivering the culture solution to the culture chamber 2 and the speed of the nutrient solution delivery unit 11 for delivering the nutrient solution to the second exchange chamber 303 are controlled according to the detected concentration and/or the determined growth condition of the culture.

EXAMPLE six

As shown in fig. 13, the present embodiment discloses an in vitro life support perfusion culture system, which includes a premixing unit 1, a culture unit, an exchange unit 3, a fluid infusion unit 11, a collection unit 4, and a power unit 5.

The in vitro life support perfusion culture system in this embodiment has substantially the same structure as the in vitro life support perfusion culture system in the first embodiment, except that the first exchange chamber 302 directly supplies the culture solution to the culture chamber 2 without directly supplying the culture solution to the premix chamber 101 in a reverse flow manner, and the power unit 5 is provided between the first exchange chamber 302 and the culture chamber 2, and the culture solution in the second exchange chamber 302 is transferred into the culture chamber 2 by using the power unit 5. At this time, the premix chamber 101 also continuously feeds the culture medium to the culture chamber 2, and the gas necessary for the growth of the culture is continuously fed into the culture chamber 2.

It should be understood that the above-described embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. An in vitro life support perfusion culture system, comprising:

a premixing unit (1) comprising a premixing chamber (101), wherein nutrient solution and gas are added into the premixing chamber (101), and the nutrient solution and the gas are mixed in the premixing chamber (101) to form culture solution;

a culture unit comprising a culture chamber (2), wherein the culture chamber (2) is communicated with the premixing chamber (101), culture fluid in the premixing chamber (101) can be conveyed into the culture chamber (2), and the culture chamber (2) is used for containing a culture and culturing the culture;

an exchange unit (3) comprising an exchange chamber and a membrane module (301), wherein the exchange chamber comprises a first exchange cavity (302) and a second exchange cavity (303), the membrane module (301) is arranged between the first exchange cavity (302) and the second exchange cavity (303) and is communicated through the membrane module (301), the culture chamber (2) and the premixing chamber (101) are both communicated with the first exchange cavity (302), the first exchange cavity (302) can receive culture fluid flowing out of the culture chamber (2) and can convey the culture fluid to the premixing chamber (101) and/or the culture chamber (2), and the membrane module (301) is used for intercepting and/or permeating partial components in the culture fluid;

a liquid supplementing unit (11) communicated with the second exchange cavity (303) and used for delivering nutrient solution to the second exchange cavity (303), wherein the liquid supplementing unit (11) can directionally deliver one or more components required by the culture to the second exchange cavity (303), and at least part of components in the nutrient solution can permeate into the first exchange cavity (302) through the membrane component (301).

2. The in vitro life support perfusion culture system of claim 1, wherein the first exchange chamber (302) is provided with a first interface and a second interface, the first interface is in communication with the culture chamber (2), and the second interface is in communication with the premix chamber (101) and/or the culture chamber (2);

the culture solution in the culture chamber (2) can flow to the first exchange cavity (302) through the first interface, and the culture solution in the first exchange cavity (302) can flow to the premix chamber (101) and/or the culture chamber (2) through the second interface.

3. The in vitro life support perfusion culture system of claim 2, further comprising a power unit (5), wherein the power unit (5) is arranged between the pre-mixing chamber (101) and the culture chamber (2), wherein the power unit (5) is arranged between the culture chamber (2) and the first interface, and wherein the power unit (5) is arranged between the pre-mixing chamber (101) and the second interface and/or between the culture chamber (2) and the second interface.

4. The in vitro life support perfusion culture system of claim 1, further comprising a collection unit (4), the collection unit (4) being in communication with the second exchange chamber (303) for collecting liquid within the second exchange chamber (303).

5. The in vitro life support perfusion culture system of claim 4, wherein at least one of the pre-mixing unit (1), the culture chamber (2), the exchange unit (3) and the collection unit (4) is a disposable consumable.

6. The in vitro life support perfusion culture system of claim 4, wherein the culture chamber (2) is multiple, and the multiple culture chambers (2) are all communicated with the premixing chamber (101);

one exchange unit (3) is connected to each of the culture chambers (2), or all of the culture chambers (2) are connected to one exchange unit.

7. The in vitro life support perfusion culture system of claim 1, further comprising:

the temperature control unit (6) comprises a refrigeration assembly and a temperature control module, the refrigeration assembly is electrically connected with the temperature control module, the temperature control module is used for controlling the refrigeration assembly to cool the culture chamber (2) to a first preset temperature, and the first preset temperature is the liquefaction temperature of the culture support structure;

a sampling port (202) is arranged on the culture chamber (2), and the sampling port (202) is used for taking out the culture after the culture supporting structure is liquefied.

8. The in vitro life support perfusion culture system of claim 1, wherein the premixing unit (1) further comprises a pH detector (102), a dissolved oxygen detector (103), and a signal detector, wherein the pH detector (102) and the dissolved oxygen detector (103) are both disposed in the premixing chamber (101), and the signal detector can sense signals fed back by the pH detector (102) and the dissolved oxygen detector (103) to obtain the pH value and the dissolved oxygen amount of the culture fluid in the premixing chamber (101).

9. The in vitro life-supporting perfusion culture system of claim 8, wherein the pH detector (102) is a pH electrode pad, and the dissolved oxygen detector (103) is an dissolved oxygen electrode pad;

the pH electrode plate is arranged on the inner wall of the premixing chamber (101) or in the premixing chamber (101), and the dissolved oxygen detector (103) is arranged on the inner wall of the premixing chamber (101) or is immersed in liquid in the premixing chamber (101).

10. The in vitro life support perfusion culture system of claim 1, wherein the pre-mixing unit (1) further comprises a gas pre-mixing control unit (105) in communication with the pre-mixing chamber (101) for delivering gas into the pre-mixing chamber (101).

11. The in vitro life support perfusion culture system of claim 10, wherein the number of the premixing chambers (101) is multiple, the number of the gas premixing control units (105) is one, and the gas premixing control units are simultaneously communicated with the plurality of premixing chambers (101) to control the gas concentration in the plurality of premixing chambers (101).

12. The in vitro life support perfusion culture system of claim 1, further comprising: a sterility control module (9) comprising a sterile working chamber (901), a filter assembly (903) and a sterilization assembly (904), at least the culture chamber (2) being arranged within the sterile working chamber (901), the filter assembly (903) being adapted to filter the gas passing into the sterile working chamber (901), the sterilization assembly (904) being adapted to sterilize the sterile working chamber (901).

13. A method for controlling an in vitro life support perfusion culture system according to any one of claims 1-12, comprising the steps of:

detecting the concentration of the components required by the culture in the culture solution in the culture chamber (2) and/or determining the growth condition of the culture, and controlling the speed of the premix chamber (101) for delivering the culture solution to the culture chamber and the speed of the liquid supplementing unit (11) for delivering the nutrient solution to the second exchange cavity (303) according to the detected concentration and/or the determined growth condition of the culture.