BHK21 cell serum-free culture medium

Abstract

The invention discloses a BHK21 cell serum-free culture medium, including glucose, galactose, a sericin hydrolysate, corn flour agar, amino acids, minerals, a nonionic surfactant, a fatty acid ester complex agent, vitamins, ethanolamine, an antioxidant and a pH buffer. Cells grow in a semi-adherent and semi-suspended state when the cells are statically cultured in the medium, and the cell survivalrate is 95% or more. The culture medium has no animal-derived components, and improves the biosafety of a final product. The culture medium supports cell growth and also supports a virus proliferation process in a later stage of culture, and a higher virus titer is generated. The preparation method is simple, the price is lower, and the culture medium has a scale-up production and application prospect.

Description

technical field [0001] The invention relates to the technical field of medium preparation, in particular to a BHK 21 Serum-free medium for cells. Background technique [0002] The host cells currently used for recombinant viral vector amplification, recombinant protein expression or vaccine scale preparation are basically derived from mammalian cells, such as young hamster kidney cells (BHK 21 ), these cells usually need to add a certain amount (5-10%) of bovine serum in the traditional culture process, which contains growth factors, hormones, carrier proteins, adhesion factors, trace elements and other nutrients required for cell growth , can promote cell growth. However, in large-scale production and practical application, the addition of serum has the following disadvantages: there is no commercially available fully chemically synthesized product, and the chemical composition of animal hydrolyzed milk protein is not fixed, so the quality of the medium cannot be guarante...

AI Technical Summary

Problems solved by technology

However, in large-scale production and practical application, the addition of serum has the following disadvantages: there is no commercially available fully chemically synthesized product, and the chemical composition of animal hydrolyzed milk protein is not fixed, so the quality of the medium cannot be guaranteed. thus making BHK 21 The quality of the vaccine synthesized by cells cannot be guaranteed; moreover, the shelf life of animal-derived hydrolyzed milk protein is generally short, and the storage environment needs to be strictly controlled, otherwise it is easy to deteriorate; at the same time, the three-in-one preparation used in the medium The process of the method is more loaded down with trivial details, and production efficiency is lower

[0003] In order to overcome the above-mentioned defects brought about by the addition of serum, many scientific research institutes or biopharmaceutical companies have successively developed several serum-free media since the 1980s, but there are also shortcomings: 1. In the serum-free media It still contains animal protein source components, such as bovine serum albumin (BSA), insulin, transferrin, etc., and there are still problems that may occur in the application of serum. In addition, it is expensive and generally only suitable for small-scale cultivation in laboratories; 2. The currently developed serum-free medium has high specificity for cells, and usually a medium can only be suitable for the growth of one cell line, but not for other cell lines;

Images