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Siphon type cultivation method

A medium and culture medium technology, applied in general culture methods, tissue culture, cell culture supports/coatings, etc., can solve the problems of upper limit of cell density and increased cost

Inactive Publication Date: 2019-03-15
UBE IND LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the volume of the medium in which the microcarriers are dispersed needs to be stirred and diffused as fully as possible, so there is an upper limit to the density of cells that can be cultured
In addition, in order to separate the microcarriers from the culture medium, it is necessary to separate them with a filter capable of classifying fine particles, which also causes an increase in cost.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0227] Cell Culture Method Using Porous Polyimide Membrane Using Siphon Reactor (Part 1)

[0228] The acclimatized and suspended cells of CHO-DP12 cells (ATCC CRL-12445) producing anti-human IL-8 antibody were suspended and cultured in medium (BalanCD (trademark) CHO GROWTH A), and the culture was continued until the number of viable cells per 1 ml was 3.9×10 6 . Inject 12ml of the above-mentioned suspension culture solution into a petri dish with a diameter of 10cm, and then use a nylon mesh (30#, mesh 547μm) to form a coat (shell), and add a certain amount (20cm per module) to the petri dish. 2 12 modules of the sterilized polyimide porous membrane. After wetting the module with the cell suspension, the 2 Let stand overnight in the incubator.

[0229] On day 2, remove the module and place in figure 1 The shown siphon reactor was provided with 10 modules in the step section, and 350 ml of medium (IMDM containing 2% FBS) was stored in the reservoir, and the medium was cir...

Embodiment 2

[0231] Cell Culture Method Using Porous Polyimide Membrane Using Siphon Reactor (Part 2)

[0232] The acclimatized and suspended cells of CHO-DP12 cells (ATCC CRL-12445) producing anti-human IL-8 antibody were suspended and cultured in medium (BalanCD (trademark) CHO GROWTH A), and the culture was continued until the number of viable cells per 1 ml was 9.9×10 6 . Load 10 modules into an oxygen permeable bag for shaking culture in CO 2 Shaking culture was carried out in the incubator overnight. On the second day, the module was taken out from the shaking bag, and under the same conditions as in Example 1, the cells were cultured in a siphon reactor. 350 ml of a culture medium (KBM270 manufactured by Cogent Bio) was stored in the reservoir, and the culture medium was circulated at a rate of 60 ml / min via a tube pump. After 4 days, the culture ended, at which time it was observed that the cell density was 1.0×10 5 cells / cm 2 , the total number of cells is 2.1×10 7 cells. ...

Embodiment 3

[0234] Metal modules and glass heat-resistant siphon reactors

[0235] The heat resistance of the polyimide porous membrane should be utilized to the maximum, and the sterilization operation should be completed by simple overall dry heat sterilization, and the outer shell, the lining and the polyimide porous membrane made of stainless steel mesh should be produced Consists of metal modules (see image 3 One embodiment of (B)). Specifically, a 1 cm × 1 cm porous polyimide membrane and a stainless steel mesh having the same area as the porous polyimide membrane (called "interlining", not shown) are laminated (according to Porous Polyimide 3 Sheets, 1 sheet of lining, 4 sheets of porous polyimide, 1 sheet of lining, and 3 sheets of porous polyimide were stacked in this order), and the laminated product was sealed with a stainless steel mesh casing to produce a metal module. Work can be performed non-sterile in an open space.

[0236] The heat-resistant siphon reactor made of g...

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Abstract

The present invention provides a cell cultivation device comprising a polymer porous film, and a cell cultivation method using said cell cultivation device. Provided is a cell cultivation device thathas a siphon structure or a shishi-odoshi structure, said cell cultivation device comprising: a polymer porous film; a cell cultivation part that accommodates the polymer porous film; a liquid reservoir part that has the cell cultivation part stored in an interior thereof; a culture medium supply means that is positioned at an upper section of the liquid reservoir part; a culture medium dischargemeans that is positioned at a lower section of the liquid reservoir part; and a culture medium recovery means that recovers the culture medium that is collected in the liquid reservoir part and is intermittently expelled.

Description

technical field [0001] The present invention relates to a cell culture device provided with a polymer porous membrane. In addition, it relates to a cell culture method using a cell culture device provided with a polymer porous membrane. Background technique [0002] In recent years, proteins such as enzymes, hormones, antibodies, cytokines, viruses (viral proteins) for therapeutics and vaccines have been industrially produced using cultured cells. However, such protein production techniques are costly, which increases medical costs. Therefore, in order to significantly reduce costs, a technique for culturing cells at a high density and an innovative technique for increasing protein production are being sought. [0003] As protein-producing cells, anchorage-dependent adherent cells that adhere to a culture substrate are sometimes used. Since such cells proliferate in an anchorage-dependent manner, they must be cultured by attaching them to the surface of a culture dish, cu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C12N1/00
CPCC12N1/00C12M29/12C12M25/02C08G73/1007C08G73/1067C08L79/08C08L81/06C12M23/44C12M25/14C12M27/10C12M29/06C12N5/0068C12N2533/30C12M23/06C12M23/34C12M29/04C12M1/00
Inventor 萩原昌彦布施新作
Owner UBE IND LTD
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