CN103614296B - Double-chamber three-dimensional pouring bioreactor system - Google Patents
Double-chamber three-dimensional pouring bioreactor system Download PDFInfo
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Abstract
The invention discloses a double-chamber three-dimensional pouring bioreactor system which comprises a bioreactor, wherein the bioreactor is internally provided with a three-dimensional bracket and is connected with a pouring device. The bioreactor comprises a first chamber filled with common nutritional nutrient liquor and a second chamber filled with inductive culture factors. A dialysis membrane assembly through which components having molecular weight being less than the molecular weight of inductive culture factors in the inductive culture factors can pass is arranged between the first chamber and the second chamber. The three-dimensional bracket is located in the second chamber. According to the double-chamber structural design of the bioreactor disclosed by the invention, the common nutritional nutrient liquor and the inductive culture factors in the two chambers are independently exchanged, and the inductive culture factors can be preserved while the common nutritional nutrient liquor is changed, so that the use level of the inductive culture factor is greatly reduced, the effective utilization ratio of the inductive culture factors is improved, and the cost is lowered.
Description
Technical field
The invention belongs to biotechnology apparatus field, especially relate to the two-chamber dimensional perfusion bioreactor system of cell Growth and Differentiation in three-dimensional rack.
Background technology
At present, in life science field and medical field, cell cultures mode has multiple, substantially can be classified as two dimension and dimensional culture mode by the dimension classification of cultivating.In two dimension training method, as culturing bottle, the culture dish of glass or plastics, need to provide a plane for cell attachment growth, by providing cell better growing environment to the classification of planar materials and the modification of contact surface, same can the growth conditions of observation of cell very easily by inverted microscope and simple microscope.But normal cell is all grow in three-dimensional environment in human body and animal body, some cell cannot adhere to any planar growth, therefore in vitro for cell provides a kind of environment of three-dimensional to carry out cultivating the normal physiological condition more meeting cell, the corresponding process of growth in better analogue body is contributed to.
Dimensional culture has various ways in current technical field, as: (1) cultivates on the basis of plane in two dimension, side adds with one deck collagen matrices composition in the plane, cell is grown in the three-dimensional environment of matrix, also make three-dimensional rack by artificial material, cultivate in conjunction with perfusion system; (2) by the swing of revolving oar and the powered rotation of different shape, cell suspension is grown in the three-dimensional environment cultivating liquid, also have by some artificial materials by single or multiple cell micro-encapsulation, for cell provides attachment surface to contact with intercellular; (3) manufacture a kind of weightless environment by centrifugal principle, make to grow to all directions in cell energy three-dimensional space.
The mode of cultivating is carried out in conjunction with filling system by three-dimensional rack in above-mentioned three-dimensional cell cultivation mode, if notification number is " a kind of rotary pouring type bioreactor system " disclosed in the patent document of CN 1288235C, it comprises bio-reactor, this bio-reactor comprises base, base is provided with reaction vessel, be filled with nutrient solution in this reaction vessel and be installed with three-dimensional rack, nutrient solution is injected reaction vessel by filling system by this reaction vessel, and the gaseous interchange inside and outside bio-reactor is carried out by semi-permeable membranes in these reaction vessel two ends.This rotary pouring type bioreactor system, cell culture environment can well be improved, promote cell being more evenly distributed on three-dimensional rack, promote that oxygen and nutritive substance are carried to internal stent, keep the activity of internal stent cell and the performance of function, gas real-time exchange inside and outside realization response device, ensure that the content of oxygen and carbonic acid gas in nutrient solution is basicly stable, maintain physiological pH value, for cellular metabolism and Function provide more favourable microenvironment.Nutrient solution in above-mentioned bio-reactor is in order to promote the growth and differ entiation of cell, generally comprise ordinary nutritional nutrient solution and the inducing culture factor, ordinary nutritional nutrient solution generally comprises basic nutrient as growth and the propagation to maintain cell such as sugar, amino acid, trace element, also comprise damping fluid to regulate suitable physiological PH value, the inducing culture factor then as Growth of Cells differentiation key component play a role, but because of its more difficult acquisition and yield poorly down and exception expensive.Because above-mentioned bio-reactor only has a reaction vessel, then grown cultures liquid and the inducing culture factor all must be blended in same reaction vessel, the required inducing culture factor consumption once cultivated is very large; Inexpensively, for meeting Growth of Cells, nutrient solution needs regular replacing to ordinary nutritional nutrient solution, and at every turn along with the replacing of ordinary nutritional nutrient solution, the inducing culture factor also runs off thereupon, so cultivates, and the cost spent is higher.
Summary of the invention
In order to overcome the deficiencies in the prior art, the invention provides a kind of two-chamber dimensional perfusion bioreactor system, this two-chamber dimensional perfusion bioreactor system improves the utilization ratio of the inducing culture factor greatly by the design of double cavity structure and dialysis membrane assembly, reduce toxigenic capacity, cultivate more economical material benefit compared with dimensional perfusion formula of the prior art.
To achieve these goals, the technical solution used in the present invention is: a kind of two-chamber dimensional perfusion bioreactor system, comprise bio-reactor, three-dimensional rack is provided with in bio-reactor, bio-reactor is connected with device for casting, it is characterized in that: described bio-reactor comprises first cavity volume of filling for ordinary nutritional nutrient solution, and for the second cavity volume that the inducing culture factor is filled, the dialysis membrane assembly that the composition that can be less than inducing cytokine in the inducing culture factor for molecular weight passes through is provided with between first cavity volume and the second cavity volume, described three-dimensional rack is arranged in described second cavity volume.
In said structure, the design of double cavity structure makes ordinary nutritional nutrient solution and the inducing culture factor be placed in two chambeies, and by the dialysis membrane assembly between two-chamber, the nutrient needed for growth and proliferation of cell can being exchanged in two chambeies at any time, the macromolecular substance such as the inducing cytokine in the inducing culture factor are then isolated in the second cavity volume; The inducing culture factor can be retained when changing ordinary nutritional nutrient solution, greatly reducing the consumption of inducing cytokine, improve its effective rate of utilization, greatly reduce toxigenic capacity.
As the further setting of the present invention, micropore or microchannel is provided with in described three-dimensional rack, described micropore or microchannel converge formation two inlet and outlet pipings and are respectively artery liquid-inlet pipe and vein drain pipe, and described artery liquid-inlet pipe and vein drain pipe are connected to form the internal liquid circulation mechanism of three-dimensional rack respectively with described device for casting.
In said structure, three-dimensional rack can be provided with the micro-or microchannel of natural or artificial three-dimensional, and the internal liquid circulation mechanism of a similar human vas circulation is finally formed by this micropore or microchannel, three-dimensional rack can be full of the full three-dimensional structure of formation one, cell is made to be attached on three-dimensional rack better, the cell in each corner of internal stent is made to obtain good gas and metabolic substd exchange, when using organ decellularization support and some have the man-made support of good biocompatibility, the suitable tissue of shape and organ can be formed in the cell cultures later stage, and to be transplanted in body on corresponding blood vessel by arteriovenous liquid in-out Guan Zhi, above-mentioned artery liquid-inlet pipe and vein drain pipe refer to that the liquid be similar in human vas is flow to by artery, the sanguimotor install pipeline that vein flows out.
As the further setting of the present invention, described internal liquid circulation mechanism comprise respectively with described artery liquid-inlet pipe and the needle assembly that is connected with described vein drain pipe, the other end that described needle assembly is connected with described artery liquid-inlet pipe or vein drain pipe is relatively connected with described device for casting, described needle assembly comprises at least two syringe needles, be respectively the first syringe needle and the second syringe needle, the afterbody of the first syringe needle is connected in series by the head head and the tail of silica gel head and described second syringe needle.
In said structure, needle assembly is connected by two syringe needles, and the silica gel head of two needle support has one end that certain elasticity makes this first syringe needle be connected with artery liquid-inlet pipe and has better oscillatory, is convenient to installation three-dimensional rack and the installation with needle assembly; Arranging of this silica gel head can play the effect that Anti-bubble enters the second cavity volume in addition, after this silica gel head is full of liquid, the stopping property connected between two syringe needles is good, and after the connection of the second syringe needle and device for casting disconnects, if do not have the setting of silica gel head, the gas in air can enter in the second cavity volume thereupon, time serious, the pipe vein system system in three-dimensional rack can be blocked, affect the cultivation of cell.
As the further setting of the present invention, described second cavity volume sidewall has the through hole passed for described needle assembly, described second cavity volume sidewall is provided with the support chamber of protruding outwardly, the head of described first syringe needle is arranged in described second cavity volume through described through hole, and the afterbody of described first syringe needle is fixedly arranged on described second cavity volume sidewall by described support chamber.
In said structure, needle assembly only has its head to be arranged in the second cavity volume, and other parts are installed in outside the second cavity volume by the support chamber that is fixedly arranged on the second cavity volume sidewall, increase in the second cavity volume can operating space, improve flexibility of operation, as the operation etc. of fixed support, connecting needle assembly and support.
As the further setting of the present invention, two sidewalls that described second cavity volume is relative are respectively equipped with outer circulation import and outer circulation outlet, and described outer circulation import and outer circulation outlet are connected to form the outside liquid cycling mechanism of one-way flow respectively with described device for casting.
In said structure, this outside liquid cycling mechanism refers to and is positioned at three-dimensional rack outside, and be positioned at the fluid circulation mechanism of the second cavity volume, this circulation is the cycling mechanism being positioned at three-dimensional rack outside of one-way flow, this outside liquid cycling mechanism can reduce the situation of cell adhesion on the second cavity volume inwall or bottom surface, make cell adhered thereto be brought in three-dimensional rack by the circulation of this outside liquid as far as possible, allow cell cultivate in three-dimensional rack; Inner-outer circulation with the use of, effect is better; In addition, the nutrient solution circulation at interval can also be carried out by controlling device for casting, making the cell in three-dimensional rack more easily can be affixed in three-dimensional rack and can not be flushed away.
As the further setting of the present invention, two sidewalls of one end that described second cavity volume is connected with described first cavity volume offer draw-in groove, described dialysis membrane assembly is embedded in draw-in groove, the Mierocrystalline cellulose semi-permeable membranes that described dialysis membrane assembly comprises the first clamping plate, the second clamping plate and is attached between the first clamping plate and the second clamping plate.
In said structure, rectangular communicating passage is provided with between first cavity volume and the second cavity volume, the both sides sidewall of this communicating passage offers corresponding draw-in groove respectively in its vertical direction, this dialysis membrane assembly is embedded at the effect playing isolation first cavity volume and the second cavity volume in this communicating passage by draw-in groove, easy for installation, be convenient to change simultaneously; Above-mentioned first clamping plate and the second clamping plate are that the clamping plate of band groove structure are arranged, and semi-permeable membranes to be located between two clamping plate and to be arranged in groove, the good sealing effect of this vibrational power flow and dialysis buffer action is good.
As the further setting of the present invention, in described second cavity volume, be embedded with the partition blocking described first cavity volume and be communicated with the second cavity volume towards one end of described dialysis membrane assembly.
In said structure, this partition can completely cut off two cavity volumes completely, and what facilitate two cavity volumes independently changes liquid.
As the further setting of the present invention, described bio-reactor also comprises the bulb apparatus that degass, its bubble assembly described comprises liquid storage pipe, the lower end of liquid storage pipe is connected with described device for casting, the upper end of liquid storage pipe is connected with negative pressure suction device, and the sidewall of liquid storage pipe is provided with the draining hole be communicated with described second cavity volume.
In said structure, the negative pressure suction device degassed in bulb apparatus can adopt the syringe removing syringe needle, negative pressure suction device can will have the bubble removal of generation in the second cavity volume, guarantee that the liquid entered in the second cavity volume does not have bubble, guarantee that the pipe vein system system in three-dimensional rack can not be blocked by bubble.
As the further setting of the present invention, the floorage of described second cavity volume is 1/3 of the first cavity volume, and described second cavity volume height is the half of described first cavity volume height.
In said structure, the vertical height of the second cavity volume is only the half of the first cavity volume; Bossed window is also established in the top of above-mentioned second cavity volume, and window is covered with transparent cover body; The present invention is by reducing the opening design of vertical height and the second cavity volume window, less height degree to be very easy in the second cavity volume by the cell growth condition in stereomicroscope observation three-dimensional rack in the operation relevant to three-dimensional rack and three-dimensional rack cell cultivation process, because can meet the requirement that stereoscopic microscope must be very near to viewing distance.
As the further setting of the present invention, above-mentioned bio-reactor is that transparent material is made.The transparent design of two cavity volumes, the light facilitating bottom is shining into requirement, also facilitates fluorescence real time imagery simultaneously and observes.
Adopt such scheme, the present invention can allow cell in system, obtain good nutrient and gaseous interchange by two-chamber dimensional perfusion bioreactor system, refuse can well be discharged, and cell can better growing multiplication and cell for the purpose of being divided under the effect of ordinary nutritional nutrient solution and the inducing culture factor; And the double cavity structure design of bio-reactor can make the ordinary nutritional nutrient solution in two cavity volumes and the inducing culture factor independently change liquid, the inducing culture factor can be retained while replacing ordinary nutritional nutrient solution, greatly reduce the consumption of inducing cytokine, improve its effective rate of utilization, reduce costs.
Below in conjunction with accompanying drawing, the invention will be further described.
Accompanying drawing explanation
Accompanying drawing 1 is specific embodiment of the invention structural representation;
Accompanying drawing 2 is specific embodiment of the invention structural front view;
Accompanying drawing 3 is the A-A sectional view of specific embodiment of the invention accompanying drawing 2;
Accompanying drawing 4 is the structure sectional view of the bio-reactor of the specific embodiment of the invention;
Accompanying drawing 5 is specific embodiment of the invention structure right view;
Accompanying drawing 6 is specific embodiment of the invention structure left view;
Accompanying drawing 7 is the structure sectional view of specific embodiment of the invention dialysis membrane assembly;
The structural representation of accompanying drawing 8 specific embodiment of the invention needle assembly;
Accompanying drawing 9 is the internal structure schematic diagram of the communicating passage in specific embodiment of the invention accompanying drawing 4;
Accompanying drawing 10 is the another kind of internal structure schematic diagram of the communicating passage in specific embodiment of the invention accompanying drawing 4.
Embodiment
Specific embodiments of the invention are two-chamber dimensional perfusion bioreactor system as shown in figs 1-9, comprise bio-reactor 1, three-dimensional rack 2 is provided with in bio-reactor 1, bio-reactor 1 is connected with device for casting, the first cavity volume 11 that bio-reactor comprises base 3, fills for ordinary nutritional nutrient solution, and for the second cavity volume 12 that the inducing culture factor is filled, first cavity volume 11 and the second cavity volume 12 are positioned on base 3, and base 3 plays the effect of the peristaltic pump of the power resources of support and fixed cycles device for casting; Be provided with the dialysis membrane assembly 4 that the composition that can be less than inducing cytokine in the inducing culture factor for molecular weight passes through between first cavity volume 11 and the second cavity volume 12, three-dimensional rack 2 is arranged in the second cavity volume 12.The design of double cavity structure makes ordinary nutritional nutrient solution and the inducing culture factor be placed in two chambeies, and by the dialysis membrane assembly 4 between two-chamber, the nutrient needed for growth and proliferation of cell can being exchanged in two chambeies at any time, the macromolecular substance such as the inducing cytokine in the inducing culture factor are then isolated in the second cavity volume 12; The inducing culture factor can be retained when changing ordinary nutritional nutrient solution, greatly reducing the consumption of inducing cytokine, improve its effective rate of utilization, greatly reduce toxigenic capacity.
Artificial or natural three-dimensional micropore or microchannel is provided with in above-mentioned three-dimensional rack 2, micropore or microchannel converge formation two inlet and outlet pipings and are respectively artery liquid-inlet pipe and vein drain pipe, and artery liquid-inlet pipe and vein drain pipe are connected to form the internal liquid circulation mechanism of three-dimensional rack 4 respectively with device for casting.The range of choice of above-mentioned three-dimensional rack 2 is very wide, can be the decellularization support of various histoorgan as liver, lungs, heart, kidney, Acellularized valve is by removing organ and in-house original cell, retains the three-dimensional rack 2 that extracellular matrix skeleton is formed.This three-dimensional rack 2 has the structure that inner vascular canal etc. meets normal physiological, by being connected into above-mentioned internal liquid circulation mechanism section, just can Growth of Cells three-dimensional environment in analogue body, and reach the growth and differ entiation effect of promotion stem cell.Also can select some thicker synthetic supports, as long as possess the circulation path of above-mentioned similar sanguimotor turnover, all can apply, same organ 3D print carriage also can be applied.
In said structure, the internal liquid circulation mechanism of a similar human vas circulation is formed in three-dimensional rack 2, three-dimensional rack 2 can be full of the full three-dimensional structure of formation one, cell can be attached on three-dimensional rack 2 better, the cell in three-dimensional rack 2 each corner inner is made to obtain good gas and metabolic substd exchange, when using organ decellularization support and some have the man-made support of good biocompatibility, the suitable tissue of shape and organ can be formed in the cell cultures later stage, and to be transplanted in body on corresponding blood vessel by arteriovenous liquid in-out Guan Zhi, above-mentioned artery liquid-inlet pipe and vein drain pipe refer to that the liquid be similar in human vas is flow to by artery, the sanguimotor install pipeline that vein flows out.
Above-mentioned device for casting comprises peristaltic pump 51 and pump line 52, the mouth of pipe deck that the base 3 of above-mentioned bio-reactor 1 is provided with corresponding peristaltic pump deck and coordinates with pump line 52, is installed on peristaltic pump 51 on base 3, forms an entirety, coordinate compact, reduce and take up an area space.
As shown in Fig. 3,8, above-mentioned internal liquid circulation mechanism comprise respectively with artery liquid-inlet pipe and the needle assembly 6 that is connected with vein drain pipe, the other end that needle assembly 6 is connected with artery liquid-inlet pipe or vein drain pipe is relatively connected with pump line 52, needle assembly 6 comprises two syringe needles, the afterbody being respectively the first syringe needle 61 and the second syringe needle 62, first syringe needle 61 is connected in series by the head head and the tail of silica gel head 63 and the second syringe needle 62.Needle assembly 6 is connected by two syringe needles, and the silica gel head 63 between two syringe needles has one end that certain elasticity makes this first syringe needle 61 be connected with artery liquid-inlet pipe and has better oscillatory, is convenient to installation three-dimensional rack 2 and the installation with needle assembly 6; Arranging of this silica gel head 63 can play the effect that Anti-bubble enters the second cavity volume 12 in addition, after this silica gel head 63 is full of liquid, the stopping property connected between two syringe needles is good, and after the second syringe needle 62 disconnects with the connection of device for casting, if do not have the setting of silica gel head 63, the gas in air can enter in the second cavity volume 12 thereupon, time serious, the pipe vein system system in three-dimensional rack 2 can be blocked, affect the cultivation of cell.
Above-mentioned second cavity volume 12 sidewall has two through holes passed for two needle assemblies 6, second cavity volume 12 sidewall is provided with the support chamber 121 of protruding outwardly, this support chamber 121 by bracket around forming, the afterbody that the head of the first syringe needle 61 is arranged in the second cavity volume 12, first syringe needle 61 through through hole is fixedly arranged on the second cavity volume 12 sidewall by support chamber 121.Needle assembly 6 only has its head to be arranged in the second cavity volume 12, and other parts are installed in outside the second cavity volume 12 by the support chamber 121 being fixedly arranged on the second cavity volume 12 sidewall, increase in the second cavity volume 12 can operating space, improve flexibility of operation, as the operation etc. of fixing three-dimensional rack 2, connecting needle assembly 6 and three-dimensional rack 2.
Two sidewalls that above-mentioned second cavity volume 12 is relative are respectively equipped with and export with outer circulation import and outer circulation the interface 13 be connected, and the interface 13 of the interface 13 of outer circulation import and outer circulation outlet is connected to form the outside liquid cycling mechanism of one-way flow respectively with device for casting.This outside liquid cycling mechanism refers to and is positioned at three-dimensional rack 2 outside, and be positioned at the fluid circulation mechanism of the second cavity volume 12, this circulation is a cycling mechanism of through three-dimensional rack 2 outside of one-way flow, this outside liquid cycling mechanism can reduce the situation of cell adhesion on the second cavity volume 12 inwall or bottom surface, cell adhered thereto is brought in three-dimensional rack 2 by the circulation of this outside liquid as far as possible, allows cell cultivate in three-dimensional rack 2; Inner-outer circulation with the use of, effect is better; In addition, the circulation at interval can also be carried out by controlling device for casting, making the cell in three-dimensional rack 2 more easily can be affixed in three-dimensional rack 2 and can not be flushed away.
As Fig. 3, 4, shown in 9, rectangular communicating passage 14 is provided with between above-mentioned second cavity volume 12 and the first cavity volume 11, the both sides sidewall of this communicating passage 14 offers corresponding draw-in groove 141 respectively in its vertical direction, dialysis membrane assembly 4 is embedded in draw-in groove 141, dialysis membrane assembly 4 comprises the first clamping plate 41, second clamping plate 42, and the Mierocrystalline cellulose semi-permeable membranes 43 be attached between the first clamping plate 41 and the second clamping plate 42, general inducing cytokine is all macro-molecular protein, therefore this semi-permeable membranes 43 is generally less than 3.5kd, can arrange according to the size of the different inducing cytokine molecular weight of albumen in the inducing culture factor in specific experiment.This dialysis membrane assembly 4 is embedded in communicating passage 14 by draw-in groove 141, easy for installation, is convenient to change simultaneously; Above-mentioned first clamping plate 41 and the second clamping plate 42 are that the clamping plate of band groove 44 structure are arranged, and semi-permeable membranes 43 to be located between two clamping plate and to be arranged in groove 44, the good sealing effect of this vibrational power flow and dialysis buffer action is good.
As shown in Figure 4, the partition 7 that blocking-up first cavity volume 11 is communicated with the second cavity volume 12 is embedded with in above-mentioned second cavity volume 12 towards one end of dialysis membrane assembly 4.This partition 7 can completely cut off two cavity volumes completely, and what facilitate two cavity volumes independently changes liquid.Above-mentioned communicating passage 14 sidewall is also provided with the groove 142 inserted for this partition 7, this groove 142 leaves space between partition 7 and dialysis membrane assembly 4 at above-mentioned draw-in groove 141 towards one end of the second cavity 12, and both are for being independently installed in communicating passage 14 as shown in Figure 10.Dialysis membrane assembly 4 and the dividing plate 7 of independent installation have better sealing property.
As shown in Figure 1, above-mentioned bio-reactor 1 also comprises the bulb apparatus 8 that degass, its bubble assembly 8 comprises liquid storage pipe 81, the lower end of liquid storage pipe 81 is connected with pump line 52 with by Medical tee joint pipe, the upper end of liquid storage pipe 81 is connected with negative pressure suction device 82, and the sidewall of liquid storage pipe 81 is provided with the draining hole 811 that the needle assembly 6 that is connected with the artery liquid-inlet pipe on three-dimensional rack 2 is communicated with.The negative pressure suction device 82 degassed in bulb apparatus 8 can adopt the syringe removing syringe needle, negative pressure suction device 82 can will have the bubble removal of generation in second cavity volume 12, guarantee that the liquid entered in the second cavity volume 12 does not have bubble, guarantee that the pipe vein system system in three-dimensional rack 2 can not be blocked by bubble.
The half of the floorage of above-mentioned second cavity volume 12 to be the 1/3, second cavity volume 12 of the first cavity volume 11 be highly the first cavity volume 11 height.The vertical height of the second cavity volume 12 is only the half of the first cavity volume 11; Bossed window 122 is also established in the top of above-mentioned second cavity volume 12, window 122 is covered with transparent cover body 123; The present invention is by the vertical height of minimizing second cavity volume 12, and second the window 13 of cavity volume 12 design, less height degree to be very easy in the second cavity volume 12 by the cell growth condition in stereomicroscope observation three-dimensional rack in the operation relevant to three-dimensional rack 2 and three-dimensional rack 2 cell cultivation process, because can meet the requirement that stereoscopic microscope must be very near to viewing distance.
As shown in Figure 3, the other end that above-mentioned first cavity volume 11 is connected with the second cavity volume 12 is relatively provided with pipe connecting 111, this pipe connecting 111 is arranged with sealing-ring 114, and pipe connecting 111 has been threaded access flap 112, access flap 112 offers the standard interface 113 be connected with device for casting.Relate to the interface that is connected with device for casting or pipeline in the present invention to be standardized component and to arrange, be convenient to configuration.First cavity volume 11 is designed by the pipe connecting 111 of openable, the liquid light in cavity is changed places and injects or remove, in addition, also reduce the possibility of the bacterial contamination in culturing process.This design not only can feed liquor body also can air inlet body, combined by bubbling style gas and send out, can well avoid culture apparatus cell to be damaged by the bubble that bubbling device produces.
Above-mentioned bio-reactor 1 is made for transparent material, can be made up according to different working condition of optically transparent polystyrene and other polyethylene materials.The transparent design of two cavity volumes, the light facilitating bottom is shining into requirement, also facilitates fluorescence real time imagery simultaneously and observes.
Adopt such scheme, the present invention can allow cell in system, obtain good nutrient and gaseous interchange by two-chamber dimensional perfusion bioreactor system, refuse can well be discharged, and cell can better growing multiplication and cell for the purpose of being divided under the effect of ordinary nutritional nutrient solution and the inducing culture factor; And the double cavity structure design of bio-reactor 1 can make the ordinary nutritional nutrient solution in two cavity volumes and the inducing culture factor independently change liquid, the inducing culture factor can be retained while replacing ordinary nutritional nutrient solution, greatly reduce the consumption of inducing cytokine, improve its effective rate of utilization, reduce costs.
Claims (10)
1. a two-chamber dimensional perfusion bioreactor system, comprise bio-reactor, three-dimensional rack is provided with in bio-reactor, bio-reactor is connected with device for casting, it is characterized in that: described bio-reactor comprises first cavity volume of filling for ordinary nutritional nutrient solution, and for the second cavity volume that the inducing culture factor is filled, be provided with the dialysis membrane assembly that the composition that can be less than inducing cytokine in the inducing culture factor for molecular weight passes through between first cavity volume and the second cavity volume, described three-dimensional rack is arranged in described second cavity volume.
2. two-chamber dimensional perfusion bioreactor system according to claim 1, it is characterized in that: in described three-dimensional rack, be provided with micropore or microchannel, described micropore or microchannel converge formation two inlet and outlet pipings and are respectively artery liquid-inlet pipe and vein drain pipe, and described artery liquid-inlet pipe and vein drain pipe are connected to form the internal liquid circulation mechanism of three-dimensional rack respectively with described device for casting.
3. two-chamber dimensional perfusion bioreactor system according to claim 2, it is characterized in that: described internal liquid circulation mechanism comprise respectively with described artery liquid-inlet pipe and the needle assembly that is connected with described vein drain pipe, the other end that described needle assembly is connected with described artery liquid-inlet pipe or vein drain pipe is relatively connected with described device for casting, described needle assembly comprises at least two syringe needles, be respectively the first syringe needle and the second syringe needle, the afterbody of the first syringe needle is connected in series by the head head and the tail of silica gel head and described second syringe needle.
4. two-chamber dimensional perfusion bioreactor system according to claim 3, it is characterized in that: described second cavity volume sidewall has the through hole passed for described needle assembly, described second cavity volume sidewall is provided with the support chamber of protruding outwardly, the head of described first syringe needle is arranged in described second cavity volume through described through hole, and the afterbody of described first syringe needle is fixedly arranged on described second cavity volume sidewall by described support chamber.
5. the two-chamber dimensional perfusion bioreactor system according to claim 1-4 any one, it is characterized in that: two sidewalls that described second cavity volume is relative are respectively equipped with outer circulation import and outer circulation outlet, described outer circulation import and outer circulation outlet are connected to form the outside liquid cycling mechanism of one-way flow respectively with described device for casting.
6. two-chamber dimensional perfusion bioreactor system according to claim 1, it is characterized in that: two sidewalls of one end that described second cavity volume is connected with described first cavity volume offer draw-in groove, described dialysis membrane assembly is embedded in draw-in groove, the Mierocrystalline cellulose semi-permeable membranes that described dialysis membrane assembly comprises the first clamping plate, the second clamping plate and is attached between the first clamping plate and the second clamping plate.
7. the two-chamber dimensional perfusion bioreactor system according to claim 1 or 6, is characterized in that: be embedded with the partition blocking described first cavity volume and be communicated with the second cavity volume in described second cavity volume towards one end of described dialysis membrane assembly.
8. two-chamber dimensional perfusion bioreactor system according to claim 1, it is characterized in that: described bio-reactor also comprises the bulb apparatus that degass, its bubble assembly described comprises liquid storage pipe, the lower end of liquid storage pipe is connected with described device for casting, the upper end of liquid storage pipe is connected with negative pressure suction device, and the sidewall of liquid storage pipe is provided with the draining hole be communicated with described second cavity volume.
9. two-chamber dimensional perfusion bioreactor system according to claim 1, is characterized in that: the floorage of described second cavity volume is 1/3 of the first cavity volume, and described second cavity volume height is the half of described first cavity volume height.
10. two-chamber dimensional perfusion bioreactor system according to claim 1, is characterized in that: described bio-reactor is that transparent material is made.
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| CN105950468B (en) * | 2016-05-23 | 2018-03-02 | 西安交通大学 | A kind of bioreactor for perfusion and the bioreactor for infiltration |
| CN106010966B (en) * | 2016-06-08 | 2019-03-15 | 中山大学附属第一医院 | Multidirectional tissue engineering porous material perfusion system |
| CN106497784B (en) * | 2016-09-01 | 2019-03-22 | 奥凯（苏州）生物技术有限公司 | A kind of control pressurer system and its compress control method for realizing culture solution and waste liquid exchange |
| CN106497783B (en) * | 2016-09-01 | 2019-03-22 | 奥凯（苏州）生物技术有限公司 | A kind of dynamic circulation cell culture apparatus and cell culture processes |
| US12024696B2 (en) * | 2019-05-08 | 2024-07-02 | Molecular Devices (Austria) GmbH | System and method for organoid culture |
| CN110684642A (en) * | 2019-10-29 | 2020-01-14 | 康珞生物科技（武汉）有限公司 | Three-dimensional perfusion type cell culture instrument |
| CN112831417A (en) * | 2021-01-29 | 2021-05-25 | 上海睿钰生物科技有限公司 | In-vitro life-sustaining perfusion culture system and control method thereof |
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