CHO cell culturing method

Abstract

The invention provides a CHO cell culturing method. The CHO cell culturing method realizes maximization of cell culture density, reduces the volume of cell culture, lowers subsequent labor intensity and enables uniform and stable quality to be obtained; and production process is compact in link and easy to operate. According to the invention, liquid supplementation is carried out in the process of cell culture and virus propagation, so nutrients needed in cell proliferation are provided, the utilization rate of a nutrient solution is increased, and cost is saved. With a production process for an adenovirus vector vaccine in the invention, virus titer can reach 8 * 10<8> TCID50 / mL.

Description

technical field [0001] The invention relates to the technical field of cell engineering, in particular to a method for culturing CHO cells. Background technique [0002] Although CHO cells can be cultured on a large scale in artificially controlled bioreactors like microbial cells, their cell structure and culture characteristics are significantly different from those of microbial cells. Animal cells are much larger than microbial cells, without cell walls, low mechanical strength, sensitive to shear force, and poor adaptability to the environment; long doubling time, slow growth, susceptible to microbial contamination, antibiotics must be used for culture; oxygen demand during culture During the culture process, the cells adhere to each other and exist in the form of clusters; the primary culture cells generally degenerate and die after 50 generations of reproduction; the metabolites are biologically active, and the production cost is high, but the added value is also high....

AI Technical Summary

Problems solved by technology

[0002] Although CHO cells can be cultured on a large scale in artificially controlled bioreactors like microbial cells, their cell structure and culture characteristics are significantly different from those of microbial cells.

Animal cells are much larger than microbial cells, without cell walls, low mechanical strength, sensitive to shear force, and poor adaptability to the environment; long doubling time, slow growth, susceptible to microbial contamination, antibiotics must be used for culture; oxygen demand during culture During the culture process, the cells adhere to each other and exist in the form of clusters; the primary culture cells generally degenerate and die after 50 generations of reproduction; the metabolites are biologically active, and the production cost is high, but the added value is also high

[0003] In the prior art, experiments have confirmed that too many passages of cells have a significant impact on the growth of CHO cells in monolayer culture and the expression of adenovirus vectors, but for the growth of CHO cells in suspension culture and the expression of adenovirus vectors does not affect the expression level of CHO cells, but there are few reports on the suspension culture process of CHO cells in the prior art