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Second-order culture process technique for producing suspended MDCK cell relevant vaccine

A technology for vaccine production and suspension cells, applied in animal cells, vertebrate cells, urinary tract/kidney cells, etc., to achieve the effect of simple production process

Pending Publication Date: 2018-07-03
上海健士拜生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Up to now, there is no relevant report on the second-stage culture method of MDCK suspension cells

Method used

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  • Second-order culture process technique for producing suspended MDCK cell relevant vaccine

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Experimental program
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Effect test

Embodiment 1

[0051] Embodiment 1 avian influenza virus is tested in the secondary culture process test of MDCK suspension cell

[0052] 1. Experimental materials

[0053] Cell line: MDCK cell line derived from ATCC, independently domesticated by Jianshun Biotech, can grow in suspension. The acclimatization condition is to reduce the serum of MDCK adherent cells to less than 5% and pass on for more than 3 generations, and then carry out suspension acclimatization culture, and the acclimatization condition is 5% CO 2 , the shaker speed was 120RPM / min, and the MDCK cell lines capable of suspension growth were screened out.

[0054] Medium: CD MDCK SFM serum-free medium (JSB, DP197, Cat. No. 88022-197).

[0055] Avian influenza virus Re-6 (Valneva Company, France).

[0056] 2. Method

[0057] Include the following steps

[0058] (1) Recovery and passage of cells

[0059] (1) Preparation of culture medium: prepare liquid culture medium CD MDCK SFM 20L according to the instructions, and st...

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PUM

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Abstract

The invention relates to a second-order culture process technique for producing a suspended MDCK cell relevant vaccine. According to the second-order culture process technique, a virus capable of pathologically changing on a MDCK suspended cell is produced by taking the MDCK suspended cell as a host cell in a second-order culture virus inoculation manner, namely that the MDCK suspended cell is diluted by 1.5 times by virtue of a production culture medium when the density grows to a density of 2*10<6> / mL-20*10<6> / mL, the density of the diluted cell is 1*10<6> / mL-10*10<6> / mL, and then virus is inoculated or inoculated before dilution. The production process of the technique is simple; a culture medium is not wasted; the liquid is not replaced; and the production culture medium and a growth culture medium are same or different culture mediums and are mixed in a proportion, so that the purposes of cell growth and virus production are achieved.

Description

technical field [0001] The invention relates to the production field of suspension MDCK cell-related vaccines, in particular to the technique of cultivating suspension MDCK cells in tanks of 30 L or above and cultivating suspension MDCK cells by a second-stage culture method to produce vaccines. Background technique [0002] With the transfer of spinner bottle technology and adherent culture to suspension culture and serum-free culture, high-density cell growth and low (suitable) density inoculation have been recognized by the technology in the field of vaccine production. In order to produce qualified vaccines concisely, efficiently and at low cost, the first step is to enable the original adherent cells to grow in suspension. The way to obtain suspension growth is simple domestication, that is, through the change of medium and the change of culture method, the cells that originally adhered to the growth can grow in suspension; similarly, the method of obtaining cell suspen...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N7/00C12R1/93
CPCC12N5/0686C12N7/00C12N2710/10051C12N2720/12251C12N2750/14051C12N2760/16151
Inventor 罗顺蔡仕君赵志欣王嘉琪
Owner 上海健士拜生物科技有限公司
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