JP2011152077A - Wave type culture apparatus - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a wave type culture apparatus which can control a culture solution at a more suitable temperature than those of conventional culture apparatuses. <P>SOLUTION: The wave type culture apparatus for seesawing a culture table on which a culture container is loaded is characterized by having thermometers disposed to contact with the culture container at a plurality of places of the culture table, and a temperature-controlling means for controlling the temperature of the culture container on the basis of the measurement values of the thermometers. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a wave type culture apparatus that stirs a culture solution in a culture vessel by swinging by a seesaw motion and supplies or removes oxygen and carbon dioxide.

Conventionally, as a device for culturing animal cells, insect cells and the like in large quantities, a culture device using a culture tank having a ventilator and / or a stirrer and a glass or metal bowl-shaped cylindrical structure has been used. In such a culture device, convection is generated by the rotation of the stirring blade of the stirrer. In the case of only the aeration device, the culture solution is uniformly mixed by the rise of bubbles in the tank, and the bubbles supplied to the culture solution Through this, oxygen is supplied and carbon dioxide is supplied or removed.
In recent years, wave-type culture apparatuses have been used mainly for the purpose of reducing costs, improving operability, and reducing contamination. The wave type culture tank is used to mix the nutrient sources supplied to the culture solution and supply oxygen from the surface of the solution by rocking the culture container such as a pillow-shaped plastic bag containing the culture solution in a seesaw motion. And supply or removal of carbon dioxide gas.

One of the characteristics of this wave type culture tank is that the culturable liquid volume range is wide. Therefore, when using a vertical cylindrical structure culture tank, there is a great merit that a process can be constructed with one culture tank even in a process that requires a plurality of culture tanks. At this time, since there is a minimum cell concentration that can be proliferated, it is necessary to set the cell concentration at the start of culture to a certain level or more.
About this wave type culture apparatus, it is disclosed in the following Patent Document 1, Non-Patent Document 1, Non-Patent Document 2, and Non-Patent Document 3,

JP 2006-540323 A

Sartorius Co., Ltd. “Single Use Culture Device Biostat Cultiback RM20 / 50 Product Specification” Sartorius Co., Ltd. “Biostat Cultiback RM20 / 50 Basic -Operation Manual-” GE Healthcare Bioscience Co., Ltd. “GROWCELLS CELLCULTURE WAVEBIOREACTOR”

  By the way, the above-mentioned wave type culture apparatus of the prior art structurally deteriorates the temperature control performance when culturing in a state where the amount of the culture solution is small as compared with the case where the amount of the solution is large. This is because in a wave type culture apparatus, the temperature of the culture solution in the culture vessel is measured by a thermometer attached so as to circumscribe the center of the culture vessel. This is because the temperature of the gas in the culture vessel is measured instead of the temperature of the culture solution in the culture vessel, particularly when the amount of the culture solution is small. And if temperature control is performed based on the temperature measured in this way, naturally appropriate temperature control cannot be performed. As a result, in the wave type culture apparatus, the upper and lower limits of the culture temperature control value differ depending on whether the amount of the culture solution is large or small, and the productivity is particularly different in temperature sensitive cultures. End up.

  This invention is made | formed in view of the situation mentioned above, and aims at providing the wave type culture apparatus which can control a culture solution to a suitable temperature rather than before.

  In order to achieve the above object, in the present invention, as a first solving means related to a wave type culture apparatus, there is a wave type culture apparatus that moves a culture table on which a culture vessel is placed. A means is adopted that includes thermometers arranged so as to be in contact with the culture vessel at a plurality of locations, and temperature control means for controlling the temperature of the culture vessel based on the measured value of the thermometer.

  In the present invention, as the second solving means relating to the wave type culture apparatus, in the first solving means, a means is adopted in which the thermometer is arranged so as to be in contact with both ends and the center of the culture table.

  In the present invention, as a third solving means relating to the wave type culture apparatus, in the first or second solving means, the temperature control means calculates an average value of the measured values of the thermometers, and calculates the average A means of controlling the temperature of the culture solution based on the value is adopted.

  In the present invention, as the fourth solving means relating to the wave type culture apparatus, in the first or second solving means, the temperature control means includes a thermometer facing the culture solution in the culture vessel. Is selected according to the motion angle, and the temperature of the culture solution is controlled based on the measured value of the selected thermometer.

According to the present invention, the wave-type culture apparatus is a wave-type culture apparatus that moves a culture table on which a culture vessel is placed on a seesaw, each of which is disposed at a plurality of locations on the culture table so as to be in contact with the culture vessel And a temperature control means for controlling the temperature of the culture vessel based on the measured value of the thermometer.
As a result, in the wave type culture device, one of the thermometers can be cultured even when the culture solution is approached during the seesaw exercise, compared to a conventional thermometer placed in the center of the culture vessel. Since the culture solution in the container can be confronted, the temperature of the culture solution can be accurately measured even if the amount of the culture solution is small. In the wave type culture apparatus, the temperature control means can control the culture solution to a more appropriate temperature based on the measurement results of these thermometers.

It is a perspective view which shows the external appearance of the wave type culture apparatus A which concerns on one Embodiment of this invention. It is a perspective view of the culture stage 2 of the wave type culture apparatus A which concerns on one Embodiment of this invention. It is front sectional drawing of the thermometer 2b arrange | positioned at the culture stand 2 of the wave type culture apparatus A which concerns on one Embodiment of this invention. It is a front view which shows the seesaw motion of the culture stand 2 of the wave type culture apparatus A which concerns on one Embodiment of this invention.

  Hereinafter, an embodiment of the present invention will be described with reference to the drawings. The present embodiment relates to a wave type culture apparatus that stirs a culture solution by swinging a culture vessel containing the culture solution by a seesaw motion.

  First, a schematic configuration of the wave type culture apparatus A according to the present embodiment will be described with reference to FIG. FIG. 1 is a perspective view showing an appearance of a wave type culture apparatus A according to the present embodiment. The wave type culture apparatus A is used, for example, for the production of pharmaceutical antibodies, and cultivates cells by using microorganisms and animal / plant cells. That is, the wave type culture apparatus A is a device for culturing cells while supplying oxygen into the culture solution L, mixing the dissolved oxygen and nutrients uniformly, and further supplying or removing carbon dioxide gas. It is.

As shown in FIG. 1, the wave type culture apparatus A includes a culture vessel 1, a culture table 2, a drive unit 3, a controller 4, an intake tube 5, and an exhaust tube 6.
The culture vessel 1 is a plastic bag made of a plastic member such as polypropylene or polyethylene, and stores the culture solution L therein. The culture vessel 1 is preferably in the shape of a pillow, but may have other shapes, and the dimensions can be appropriately changed according to the user's request. Such a culture container 1 is disposable and is excellent in reducing the production cost in industrial use.

  As shown in FIG. 1, an intake tube 5 and an exhaust tube 6 are connected to the culture vessel 1, and oxygen and carbon dioxide gas are supplied through the intake tube 5, and the inside is unnecessary through the exhaust tube 6. Gas is exhausted. In addition, the culture vessel 1 can be attached to and detached from the culture table 2, and when the culture table 2 performs a seesaw motion while the culture vessel 1 is fixed to the culture table 2, a wave is generated in the culture medium L inside. And the culture solution L in the culture container 1 is stirred by the said wave, oxygen and a carbon dioxide gas are supplied or removed, and a nutrient is mixed uniformly. In addition, the said culture solution L contains the nutrient source etc. which are needed when culturing the said cell with the cell used as culture | cultivation object.

Next, the culture table 2 will be described with reference to FIG. FIG. 2 is a perspective view of the culture table 2 of the wave type culture apparatus A according to the present embodiment.
Under the control of the controller 4, the culture table 2 generates a wave in the culture solution L in the culture vessel 1 fixed to itself by performing a seesaw motion to the left and right, and agitates the culture solution L.

Then, as shown in FIG. 2, a heater 2a is embedded in the culture table 2, and a thermometer 2b is disposed. FIG. 3 is a front cross-sectional view (cross-sectional view in the front direction of FIG. 1) of the thermometer 2b arranged on the culture table 2 of the wave type culture apparatus A according to the present embodiment.
As shown in FIG. 2, the heater 2 a is embedded in one surface of the culture table 2, and controls the culture medium L to an optimum temperature for culture by heating the culture vessel 1 from below under the control of the controller 4. .

  As shown in FIG. 2, the thermometer 2b is arrange | positioned in the total three places of the both ends and center of the culture stand 2 so that the both ends and center of the culture container 1 may be contact | connected. As shown in FIG. 3, the thermometer 2b includes a temperature sensor 2b-1 at the center, and the temperature sensor 2b-1 measures the temperature of the culture medium L via the culture vessel 1.

The temperature sensor 2b-1 outputs a temperature detection signal indicating the measured temperature to the controller 4. The temperature sensor 2b-1 is provided with a heat insulating material on the lower side thereof so as not to be directly affected by the heat of the heater 2a embedded below the thermometer 2b.
Then, the controller 4 determines the temperature of the culture solution L based on the temperature detection signal input from the temperature sensor 2b-1, and controls the temperature of the heater 2a so that the culture solution L becomes an appropriate temperature.

The drive unit 3 vibrates left and right while drawing a fan shape under the control of the controller 4. The driving unit 3 repeats this left-right vibration, so that the culture table 2 continuously performs seesaw motion.
The controller 4 includes a control circuit and an operation display unit that are constituted by a CPU (Central Processing Unit), a ROM (Read Only Memory), a RAM (Random Access Memory), and the like, and a control program stored in the ROM. The entire operation of the wave type culture apparatus A is controlled based on the temperature detection signal input from the temperature sensor 2b-1 of the thermometer 2b and the operation instruction received by the operation display unit.

The operation display unit includes a touch panel and displays various screens and operation buttons for receiving operation instructions. The details of the operation of the controller 4 will be described below as the operation of the wave type culture apparatus A.
The heater 2a and the controller 4 constitute the temperature control means of this embodiment.

  The intake tube 5 is attached to the upper side of the culture vessel 1 and supplies oxygen into the culture vessel 1. Further, the exhaust tube 6 is attached to the upper side of the culture vessel 1 so as to be aligned with the intake tube 5, and discharges unnecessary gas in the culture vessel 1 to the outside. Further, for example, a 0.22 μm filter is attached to the intake tube 5 and the exhaust tube 6, and contaminants contained in the gas passing through the filter are removed.

Next, the operation of the wave type culture apparatus A according to the present embodiment configured as described above will be described in detail with reference to FIG. FIG. 4 is a front view showing the seesaw motion of the culture table 2 of the wave type culture apparatus A according to the present embodiment. In addition, the thick arrow of FIG. 4 shows the moving direction of the culture stand 2. FIG.
First, the user who wants to culture presses the culture start button on the operation display section of the controller 4 of the wave type culture apparatus A.

  The controller 4 causes the drive unit 3 to start the seesaw motion of the culture table 2 when the operation display unit receives a culture start instruction by pressing the culture start button. When the seesaw motion is started, the culture table 2 repeats the operations of (a) and (b) in FIG. In this way, the both ends of the culture table 2 are alternately lifted, so that waves such as (a) and (b) in FIG. 4 are generated and stirred in the culture solution L in the culture vessel 1.

And the controller 4 makes the heater 2a heat the culture container 1 with the start of the seesaw motion of the culture stand 2, and makes the thermometer 2b start the measurement of the temperature of the culture solution L.
In the wave type culture apparatus A, when the culture table 2 performs a seesaw motion to the left and right, the culture solution L is shifted to one side as shown in FIGS. 4 (a) and 4 (b). The thermometer 2b always faces the culture solution L in the culture container.

  When the temperature detection signal is input from the thermometer 2b, the controller 4 calculates an average value of the measurement values of the thermometers 2b based on the temperature detection signals of the three thermometers 2b. Then, the controller 4 causes the heater 2a to heat the average value using the culture vessel 1 as the temperature of the culture solution L so that the culture solution L has a preset temperature.

  In the wave type culture apparatus A according to the present embodiment as described above, the thermometer 2b is arranged on the culture table 2 so as to be in contact with both ends and the center of the culture vessel 1, and the controller 4 determines the measured values of the thermometer 2b. An average value is calculated, and the temperature of the heater 2a is controlled using the average value as the temperature of the culture medium L.

  Thereby, compared with the conventional one thermometer 2b arranged at the center of the culture vessel 1, the culture medium L is placed on one side as shown in FIGS. 4A and 4B during the seesaw motion. Even in the closed state, any one of the three thermometers 2b can oppose the culture solution L in the culture vessel 1, so that the temperature of the culture solution L can be accurately measured even if the amount of the culture solution L is small. can do. In the wave type culture apparatus A, the controller 4 calculates an average value of the measurement values based on the measurement values of the thermometer 2b, and controls the heater 2a based on the average value, thereby further increasing the culture solution. It can be controlled to an appropriate temperature.

As mentioned above, although one Embodiment of this invention was described, this invention is not limited to the said embodiment, For example, the following modifications can be considered.
(1) In the above embodiment, the controller 4 calculates the average value of the measurement values of the three thermometers 2b and controls the heater 2a based on the average value. However, the present invention is not limited to this.

  For example, since the bias of the culture medium L in the culture vessel 1 can be estimated from the motion angle of the culture table 2 during the seesaw motion of the culture table 2, the controller 4 can move the inside of the culture vessel 1 according to the motion angle of the culture table 2. The thermometer 2b facing the culture medium L may be selected, and the heater 2a may be controlled using the measured value of the selected thermometer 2b as the temperature of the culture medium L. That is, the controller 4 may switch the thermometer 2b that measures the temperature of the culture medium L according to the motion angle of the culture table 2.

(2) In the above embodiment, the three thermometers 2b are arranged at both ends and the center of the culture vessel 1, but the present invention is not limited to this.

  For example, the number of thermometers 2b is not limited to three, and two thermometers 2b may be added and the thermometers 2b may be arranged between both ends and the center. Moreover, the thermometer 2b arrange | positioned in the center of the culture container 1 may be removed, and the controller 4 may be made to control the heater 2a based on the measurement result of the thermometer 2b of the both ends of the culture container 1. FIG. That is, in the present invention, it is sufficient that at least two thermometers 2b are arranged so as to be in contact with each other.

A ... Wave type culture device, 1 ... Culture vessel, 2 ... Culture table, 2a ... Heater, 2b ... Thermometer, 3 ... Drive unit, 4 ... Controller, 5 ... Intake tube, 6 ... Exhaust tube

Claims (4)

A wave type culture apparatus for seesaw motion of a culture table on which a culture vessel is placed,
Thermometers arranged to contact the culture vessel at a plurality of locations on the culture table,
A wave type culture apparatus comprising temperature control means for controlling the temperature of the culture vessel based on a measurement value of the thermometer.   2. The wave type culture apparatus according to claim 1, wherein the thermometer is disposed so as to be in contact with both ends and a center of the culture table.   The wave type culture apparatus according to claim 1 or 2, wherein the temperature control means calculates an average value of the measured values of the thermometers and controls the temperature of the culture solution based on the average value. . The temperature control means selects a thermometer facing the culture solution in the culture vessel according to the motion angle of the culture table, and controls the temperature of the culture solution based on the measured value of the selected thermometer. Item 3. A wave type culture apparatus according to Item 1 or 2.

Images