Domestication method of full suspension culture type Marc-145 cell line

Abstract

The invention provides a domestication method of a full suspension culture type Marc-145 cell line. The domestication method comprises the following steps: (1) cultivating adherent-culture Marc-145 cells; (2) domesticating a low-serum adherent-culture Marc-145 cell line; (3) domesticating low-serum full suspension culture type Marc-145 cells; and (4) domesticating serum-free full suspension culture type Marc-145 cells. With the application of the cell domestication method disclosed by the invention, the Marc-145 cells of suspension culture can be obtained; the cells are adaptive to low-serum and serum-free full suspension culture; the density of the cells, when subjected to serum-free culture in a 7.5L reactor, can reach 8*10<6> / ml, and at least 4.4*10<10> cells can be obtained, equal to the number of cells obtained from 80-90 15L roller bottles or 18-20 40-layer cell plants; and the domestication method disclosed by the invention is significant in effect.

Description

technical field [0001] The invention relates to a domestication method of a full suspension culture type Marc-145 cell line. Background technique [0002] Marc-145 cells are epithelial cells cloned from rhesus monkey embryonic kidney cells MA-104 by Kim HS et al. in 1993. The cells can be continuously passaged and grow in an adherent manner. They are currently the research and production of porcine reproductive and respiratory syndrome virus. (Porcine reproductive and respiratory syndrome, PRRSV, commonly known as "PRRSV") and the most ideal cell line for vaccines, and can also be used for chicken infectious bronchitis virus (Infectious bronchitis virus, IBV) and porcine parvovirus (Porcine parvovirus virus, PPV) ) and other viruses in vitro propagation. Because Marc-145 grows on the wall, the cells are cultured in an adherent monolayer during virus culture and vaccine production, and the amount of cells is limited by the container (such as: cell factory, roller bottle) or ...

AI Technical Summary

Problems solved by technology

Because Marc-145 grows on the wall, the cells are cultured in an adherent monolayer during virus culture and vaccine production, and the amount of cells is limited by the container (such as: cell factory, roller bottle) or medium (such as: microcarrier) Large-scale production requires a large enough plant or expensive microcarriers and a lot of manual operations, and there are still many technical problems in the scale-up cultivation of bioreactor microcarriers, and it is still difficult to popularize and apply

At present, when Marc-145 cells are cultured in an industrial monolayer, a certain proportion of bovine serum (including fetal bovine serum or newborn bovine serum) is added to the culture medium to allow cells to grow normally. Bovine serum is not only expensive, but also comes from natural organisms. The bovine source itself may contaminate exogenous microorganisms and pathogenic factors during the collection and processing process, and there will be biosafety risks in the use

In addition, bovine serum is a natural mixture, and its specific components have not yet been fully analyzed. It will cause difficulties in the downstream purification process after being used in the production of biological products. If there is any residue, it will usually cause side reactions from the vaccinators and affect product quality.

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