JPH0838157A - Culture solution conveyer - Google Patents

Abstract

PURPOSE:To provide a culture solution conveyer capable of continuously conveying cell bodies in the culture solution together with the culture solution without damaging the cell bodies. CONSTITUTION:This culture solution conveyer involves two or more receiving tanks 1 and 6 arranged parallel to each other for temporarily storing a cell body-containing culture solution sent from a culture tank, liquid level sensors 5 for detecting the level of the culture solution in the receiving tanks, a conveying pipe 11 for conveying the culture solution from the culture tank to the receiving tanks, a discharge pipe 13 for discharging the culture solution from the receiving tank, a force feed gas inlet pipe 12 for introducing a force feed gas to the receiving tanks, gas discharge pipes 22 and 23 for discharging gas from the receiving tanks, a pressure sensor 16 for the force feed gas and control valves 3 and 8 respectively attached to the conveying pipe, the discharge pipe, the force feed gas inlet pipe and the gas discharge pipes. This conveyer involves also a controller 15 for controlling opening and closing of the respective control valves and the pressure of the force feed gas so as to continuously discharge the culture solution by repeatedly storing and discharging the culture solution into and from the respective receiving tanks in order based on signals sent from the liquid level sensors and the pressure sensor.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a culture medium transfer device,
More specifically, it relates to a culture medium transfer device capable of continuously transferring cells together with the culture medium without damaging the cells in the culture medium.

[0002]

2. Description of the Related Art In a culturing process using various types of cells using biotechnology, various types of cells that have been cultivated in a culture tank and proliferated are recycled or one-through transferred together with a culture solution, and separated after the culturing is completed. Transfer to the process is performed. Conventionally, various types of pumps have been widely used for liquid transfer, for example, a spiral (centrifugal) type, a tire flam type, a plunger type, and a roller type pump. However, when the above-mentioned pump is used to transfer the culture solution containing the cultured cells, for example, in the case of a centrifugal type, a shearing force acts between the casing and the blade due to the high speed rotation of the rotating blade, so that the culture is performed. There was a problem that the microbial cells were cut and their growth was inhibited. Such a problem remarkably occurs particularly in plant cells and animal cells, for example, filamentous forms such as spirulina and anabaena, and those having a size of several μm to several tens of μm. In addition, plunger pumps, roller pumps, tube pumps, etc. are used in small devices, but the flow rate control of the plunger pump is unstable due to the occurrence of pulsation, and the roller pump and the tube pump consume the rollers and tubes. It is hard to say that it is sufficient because maintenance work and maintenance costs are required.

[0003]

The object of the present invention is to solve the above-mentioned problems of the prior art and to transfer a culture solution continuously with the culture solution without damaging the cells in the culture solution. To provide.

[0004]

The inventions claimed in this application are as follows. (1) Two or more transfer receiving tanks arranged in parallel for temporarily storing a culture solution containing bacterial cells in a culture tank, and a liquid level sensor for detecting the culture solution level in each transfer receiving tank A transfer pipe for transferring the culture solution from the culture tank to each transfer receiving tank, a discharge pipe for discharging the culture solution in each transfer receiving tank, and a pressure-feeding gas introducing pipe for introducing a pressure-feeding gas into each transfer receiving tank, A gas discharge pipe for discharging the gas in each transfer receiving tank and a pressure sensor for detecting the pressure of the pressure-fed gas,
Based on the signals of the transfer pipes, discharge pipes, pressure-feeding gas introduction pipes and gas discharge pipes of the transfer receiving tanks and the liquid level sensors and pressure sensors, the culture solution to the transfer receiving tanks. And a control device that controls the opening and closing of each control valve and the pressure of the pressure-fed gas so that the culture solution is continuously discharged by sequentially and repeatedly storing the culture solution and discharging the culture solution from each transfer receiving tank. And a culture medium transfer device.

In the present invention, two or more transfer receiving tanks are arranged in parallel, preferably three, in order to transfer the culture solution continuously.
The discharge of the culture solution, which is provided in four units and temporarily stored in these transfer receiving tanks, is performed under static pressure by the pressurized gas without using mechanical drive. That is, the pressurized gas is sent under pressure to one tank in which the culture solution is stored to discharge the culture solution, and while the culture solution is discharged from the one tank, the culture solution is stored in another tank. Then, after the discharge of the culture solution from one tank is completed, the pumping gas is continuously supplied to the other tank to continuously discharge the culture solution.

[0006] The storage and discharge of the culture solution in each transfer tank and the introduction of the pressure-fed gas are switched by opening and closing the respective control valves provided in each transfer receiving tank. It is performed by the controller based on the signal from the surface sensor. The pressure of the pressure-fed gas is usually 1 to 1
It is 0 kg / cm ² G, but the signal detected by the pressure sensor is such that the storage and discharge of the culture solution in each transfer receiving tank are sequentially and continuously repeated according to the storage rate and the discharge rate of the culture solution. Is adjusted by the controller based on As the pressure-fed gas, various gases such as air, nitrogen gas, CO ₂ gas, and combustion exhaust gas that have been dehumidified and sterilized are used.

The culture medium transfer device of the present invention will be described in detail below with reference to the drawings. FIG. 1 is an explanatory view of a culture medium transfer device showing an embodiment of the present invention. In the figure, the culture medium transfer device includes a first transfer receiving tank 1 and a second transfer receiving tank 6 provided in parallel for temporarily storing the culture solution 20 containing the bacterial cells in the culture tank 17, and the respective tanks. Liquid level sensors 5 and 10 for detecting the culture liquid level inside, a transfer pipe 11 for transferring the culture liquid from the culture tank 17 to the tanks 1 and 6, and a discharge pipe 13 for discharging the culture liquid in each tank. A compressed gas introduction pipe 12 for introducing compressed gas into each tank, a gas discharge pipe 24 for discharging gas in each tank, a pressure sensor 16 for detecting the pressure of the compressed gas, and the transfer pipe 11
The transfer control valve 2 for the first transfer receiving tank and the transfer control valve 7 for the second transfer receiving tank, and the discharge adjusting valves 4 and 2 for the first transfer receiving tank provided in the discharge pipe 13.
Discharge control valve 9 of the transfer receiving tank and pressure-feeding gas introduction pipe 12
Of the first transfer receiving tank and the pressure transfer gas adjusting valve 8 of the second transfer receiving tank, and the gas discharge adjusting valve 22 of the first transfer receiving tank provided in the gas discharge pipe 24 and The gas discharge control valve 23 of the second transfer tank and the control valves 2, 7, 4, 9 of the first transfer receiving tank and the second transfer receiving tank based on the signals of the liquid level sensors 5, 10 and the pressure sensor 16 described above. 3, 8, 22, 23, and a control device 15 for adjusting the pressure of the pressure-fed gas.

In such a structure, first, the culture tank 17
The culture solution 20 therein is transferred to the first transfer receiving tank 1 and temporarily stored therein. The opening and closing of each control valve at this time is controlled by the controller 15 so that the transfer control valve 2 and the gas discharge control valve 22 of the first transfer receiving tank 1 are opened, the discharge control valve 3 and the pressurized gas control valve 4 are closed, and the second transfer is performed. The transfer control valve 7, the discharge control valve 8, the pressure-feed gas control valve 9 and the gas discharge control valve 23 of the receiving tank 6 are controlled to be closed.

Next, when the culture liquid level in the first transfer receiving tank 1 reaches a certain high level, the signal of the liquid level sensor 5 is sent to the control device 15, and the signal of the control device 15 causes the first
The transfer control valve 2 and the gas discharge control valve 22 of the transfer receiving tank 1 are closed, and the discharge control valve 3 and the pressure-feed gas control valve 4 are changed to open,
The transfer of the culture solution to the first transfer receiving tank 1 is stopped, the pressure-fed gas is introduced, and the culture solution is discharged from the tank 1.
At the same time, the transfer control valve 7 and the gas discharge control valve 23 of the second transfer receiving tank 6 are changed to open, and the discharge control valve 8 and the pressure-feeding gas control valve 9 are changed to closed, so that the culture from the culture tank 17 to the second transfer receiving tank 6 is started. The liquid transfer is started. The open / closed state of each control valve at this time is shown in FIG.

Further, when the culture liquid level of the first transfer receiving tank 1 reaches a certain low level, a signal from the liquid level sensor 5 is sent to the control device 15, and the signal from the control device 15 causes the open / closed state. It is changed to the opposite state. That is, the transfer control valve 2 and the gas discharge control valve 22 of the first transfer receiving tank 1 are opened, the discharge control valve 3 and the pressure-feed gas control valve 4 are closed, and the second
The transfer control valve 7 and the gas discharge control valve 23 of the transfer receiving tank 6 are closed, the discharge control valve 8 and the pressure-feeding gas 9 are changed to open, the transfer of the culture solution to the second transfer receiving tank 6 is stopped, and the pressure-feeding gas is transferred. Is introduced and the culture solution is discharged from the tank.
The culture solution is transferred from the culture tank 17 and stored in the transfer receiving tank 1. The culture solution 20 is continuously transferred from the culture tank 17 to the transfer destination by sequentially repeating the same operation.

In such a culture medium transfer device, the storage and discharge of the culture liquid in two or more transfer tanks provided in parallel are repeated in order by opening / closing each control valve and adjusting the pressure of the pressure-fed gas. Since the culture solution can be continuously transferred under the static pressure of the pressurized gas without using mechanical drive, it becomes easy to maintain the morphology of the bacterial cells in the culture solution,
Unlike the conventional method, the bacterial cells are not cut during the transfer and the growth is not inhibited. Further, such a transfer device can be applied not only to transfer of a culture solution, but also to transfer of a solid-liquid slurry containing resin particles having low strength, for example, a packing operation of a packing material for liquid chromatography into a column.

[0012]

EXAMPLES The present invention will be described below with reference to examples, but the present invention is not limited thereto. Example 1 Test bacterial cells Anabaena sp. (Cyanobacteria) at 25 ℃
3 liters of the resulting culture solution (cell concentration: 1.0 g / l) was cultivated in the modified detomer medium described in Example 1 and continuously recycled for 24 hours using the double-type culture solution transfer device shown in FIG. I went. The recycling transfer was performed by providing a circulation line connecting the discharge pipe 13 of FIG. 1 to the culture tank 17. The transfer tank has a capacity of 5 liters and a transfer capacity of 1
0 liter / min (5 liter / 30 sec / Batc
h). Air was used as the pressure-feeding gas, and the gas was introduced at a pressure of 2 kg / cm ² G. The changes in the amount of protein in the culture solution during the recycle transfer were examined, and the results are shown in Fig. 2. These changes were not observed during the transfer, indicating that the bacterial cells in the culture solution were not damaged. all right. Further, the morphology of the bacterial cells in the culture medium after 24 hours was observed with a microscope, but no cutting of the bacterial cells was observed.

Comparative Example 1 As shown in FIG. 3, a centrifugal pump (Iwaki Magnet Pump Co., Ltd., 10 l / min, 100 V, casing and impeller: carbon fiber reinforced polypropylene) was added to 3 l of the same culture solution as in Example 1. 21 was used for continuous recycle transfer for 24 hours. The change in the amount of protein in the culture solution was examined in the same manner as in Example 1, and the results are shown in FIG. 2. The amount of protein increased with the passage of time, and the bacterial cells in the culture solution were damaged. I understand. In addition, the morphology of the bacterial cells in the culture solution after 24 hours was observed with a microscope. The bacterial cells were cut, and the bacterial cells obtained in Example 1 were 1/2 to 1 times.
It was about / 4 short.

[0014]

According to the present invention, since the culture solution can be continuously transferred under static pressure without using a mechanical driving means, it is easy to maintain the morphology of the bacterial cells in the culture solution. The cells are not cut during the transfer to the cell and the growth is not inhibited.

[Brief description of drawings]

FIG. 1 is an explanatory view of a culture medium transfer device showing an embodiment of the present invention.

FIG. 2 is a graph showing changes in the amount of protein in the culture medium during transfer in Example 1 or Comparative Example 1.

FIG. 3 is an explanatory diagram of a culture medium transfer device used in Comparative Example 1.

[Explanation of symbols]

1 ... 1st transfer receiving tank, 2, 7 ... Transfer control valve, 3, 8 ...
Pressure feeding gas control valve, 4, 9 ... Discharge control valve, 5, 10 ... Liquid level sensor, 6 ... Second transfer receiving tank, 11 ... Transfer pipe, 1
2 ... Pressure-feeding gas introduction pipe, 13 ... Exhaust pipe, 15 ... Control device, 16 ... Pressure sensor, 17 ... Culture tank, 20 ... Culture liquid, 21 ... Centrifugal pump, 22, 23 ... Gas discharge control valve,
24 ... Gas exhaust pipe.

Claims (1)

[Claims] 1. Two or more transfer receiving tanks arranged in parallel for temporarily storing a culture solution containing bacterial cells in a culture tank,
A liquid level sensor for detecting the surface of the culture solution in each transfer receiving tank, a transfer pipe for transferring the culture solution from the culture tank to each transfer receiving tank, and a discharge pipe for discharging the culture solution in each transfer receiving tank. A pressure-feeding gas introducing pipe for introducing the pressure-feeding gas into each transfer-receiving tank, a gas discharge pipe for discharging the gas in each transfer-receiving tank, and a pressure sensor for detecting the pressure of the pressure-feeding gas,
Based on the signals of the transfer pipes, discharge pipes, pressure-feeding gas introduction pipes and gas discharge pipes of the transfer receiving tanks, and the signals of the liquid level sensors and pressure sensors, the culture solution to the transfer receiving tanks. And a control device that controls the opening and closing of each control valve and the pressure of the pressure-fed gas so that the culture solution is continuously discharged by sequentially and repeatedly storing the culture solution and discharging the culture solution from each transfer receiving tank. And a culture medium transfer device.