Microcarrier cell harvesting and re-inoculation amplification method based on cell reactor

Abstract

The invention provides a microcarrier cell harvesting and re-inoculation amplification method based on a cell reactor. The microcarrier cell harvesting and re-inoculation amplification method includesstep 101, filtering and collecting microcarrier cells; step 102, removing buffer solutions from culture media for balancing; step 103, performing trypsinization; step 104, separating the microcarriercells, and collecting the cells or adding the cells into a microcarrier in a next stage for inoculation and amplification. The microcarrier cell harvesting and re-inoculation amplification method hasthe advantages that large-scale microcarrier collection, controllable cell digestion, cell harvesting and microcarrier separation, and ball-to-ball amplification are realized; and the method is suitable for preparation and production of adherent cells, is applied to reagent production of vaccines, recombinant protein and biological enzymes, and is simple and economical.

Description

technical field [0001] The invention belongs to a large-scale microcarrier cell culture amplification process of a cell reactor, and in particular relates to a cell harvesting and reseeding amplification method of a cell reactor microcarrier. Background technique [0002] Microcarrier cell reactor suspension culture to prepare virus vaccine is the vaccine production mode recommended by WHO and CFDA. The volume of foreign bioreactors used for cell influenza vaccines has exceeded 6000L, and the volume of domestic microcarrier suspension culture reactors has also exceeded 300L. Although microcarrier suspension culture has the advantages of automation, easy monitoring, stable and repeatable production process, etc., the amplification process needs to digest the cells from the microcarriers, and then the cells and the old microcarriers will enter the next stage of reaction together with the new microcarriers for the next stage of culture. In the container, the image says that th...

AI Technical Summary

Problems solved by technology

In order to inoculate the next level of microcarriers, it is necessary to maintain the active state of the cells, and the rest of the microcarriers will cause the cells to be hypoxic, and the time of trypsin digestion is not well controlled to damage the cells

[0003] At present, the existing ThermoFisher company sells commercial cell collection bags for collecting microcarrier cells, which are mainly used for dozens of liters of experiments and pilot scales. The cavity of the collection bag is small, and the microcarriers are easy to clog and accumulate, and the digested cells cannot be fully released. Collection, and this collection bag is airtight, the cell digestion process needs to be controlled to prevent cell hypoxia

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