Culture solution for culturing 293T cells in serum-free suspension mode

Abstract

The invention discloses a culture solution for culturing 293T cells in a serum-free suspension mode. According to the culture solution for culturing the 293T cells in the serum-free suspension mode, ahigh-glucose DEMEM-F12 culture substrate without L-glutamine serves as a base solution, and L-alanyl-L-glutamine, vitamin C, HEPES, an insulin-like growth factor, an epidermal growth factor, lipid concentrate, human serum albumin, transferrin, trehalose, heparin sodium and Pluronic F-68 are added. The culture solution for culturing the 293T cells in the serum-free suspension mode contains clear components without any serum components, is explicit in component, stable in batch, controllable in quality, low in cost and suitable for not only routine culture in laboratories but also large-scale production culture, and has broad application prospects.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a culture medium for serum-free suspension culture of 293T cells. Background technique [0002] 293T cells are human kidney epithelial cell lines derived from 293 cells and express SV40 large T antigen, and are the main cells for packaging viruses. With the rapid development of cell therapy and gene therapy technology, especially since the successful launch of CAR-T therapy drugs, the industrialization and scale of 293T serum-free suspension culture is more urgent. [0003] Although ATCC has commercialized 293T cells to obtain a relatively stable cell line, in the actual production process, due to the large amount of 293T cells used and the excessive number of passages, the biological characteristics of the cells will change and affect the virus. packaging efficiency. At present, the conventional culture method of 293T cells mostly adopts adherence to the wall and adding serum for ...

AI Technical Summary

Problems solved by technology

[0003] Although ATCC has commercialized 293T cells to obtain a relatively stable cell line, in the actual production process, due to the large amount of 293T cells used and the excessive number of passages, the biological characteristics of the cells will change and affect the virus. packaging efficiency

At present, the conventional culture method of 293T cells mostly adopts adherence to the wall and adding serum for culture, which leads to low culture efficiency, low space utilization rate, and increased costs of instruments, consumables, labor, etc.; however, the composition of serum is complex, and the differences between batches are relatively large. Large, the test of serum quality is more complicated, the quality of the virus produced with this cell is difficult to control, and the increase of product testing items consumes a lot of manpower and material resources

However, most of the currently commercially available suspension serum-free media have problems such as low suspension acclimatization efficiency, slow cell proliferation, unstable cell characteristics, and high price.

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