Baby hamster kidney (BHK)-21 cells obtained by high-density suspension culture in low-serum and serum-free culture medium and preparation method thereof

Abstract

The invention provides BHK-21 cells obtained by high-density suspension culture in a low-serum and serum-free culture medium and a culture method thereof. The culture method comprises the following steps: 1) resuscitating frozen BHK-21 cells and performing adherent culture to obtain adherently cultured BHK-21 cells; 2) subculturing the adherently cultured BHK-21 cells in culture solution to obtain suspension-cultured BHK-21 cells and freezing the BHK-21 cells; 3) performing resuscitation culture of the suspension-cultured BHK-21 cells and performing biological property identification; and 4) performing bioreactor adaptive culture of the suspension-cultured BHK-21 cells identified to be qualified. The invention has the advantages that: a cell domesticating method provided by the invention and the obtained BHK-21 cells which can be cultured in a suspended manner save culture solution and bovine serum, are in accordance of current energy-saving, environment-protection and low-carbon concepts for the application of suspended-cultured cells into actual production saves floor area and labor investment greatly, and create considerable economic benefit and social benefit.

Description

technical field [0001] The invention belongs to the field of cell culture, and in particular relates to BHK-21 cells cultured in high-density suspension in low-serum and serum-free medium and a preparation method thereof. Background technique [0002] BHK-21 cells are Syrian hamster kidney cells, English name: Baby Hamster Kidney cell referred to as BHK-21 or BHK 21 , the cell line was established by Macpherson IA and Stoker MGP in 1961. It was derived from the kidneys of five 1-day-old hamsters of unidentified gender. Single-cell cloning was carried out in 1963. It is widely used to multiply various viruses and produce veterinary vaccines, such as foot-and-mouth disease vaccines. Because the original BHK-21 cell line is a fibroblast and grows in an anchorage-dependent manner, 10-liter and 15-liter roller bottles are used for cultured in an adherent monolayer when producing biological products. Due to the limited surface area of ​​the roller bottle, it is difficult to obtai...

AI Technical Summary

Problems solved by technology

Because the original BHK-21 cell line is a fibroblast and grows in an anchorage-dependent manner, 10-liter and 15-liter roller bottles are used for culture in an adherent monolayer when producing biological products. Due to the limited surface area of ​​the roller bottle, it is difficult to obtain enough Cells to express the corresponding viruses also require large-scale factory facilities and many manual operations

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