Suspension cell line and domestication method thereof

Abstract

The invention relates to a suspension cell line and an acclimation method thereof. The acclimation method of the suspension cell line comprises the following steps: resuscitating adherent cells in a culture medium containing 9.5-10.5% of serum, inoculating into a square bottle filled with the culture medium containing 4.5-5.5% of serum, carrying out subculture for 1-3 times, then sequentially transferring the adherent cells from the square bottle, inoculating into shaking flasks with the serum contents in the culture medium of being 3.0-3.6%, 1.8-2.2% and 0.8-1.2% sequentially, carrying out shaking flask suspension culture for 1-4 generations, then transferring into a shaking flask filled with a serum-free culture medium, and carrying out subculture for 1-3 times, so that the suspension cell line is obtained. Compared with the traditional acclimation method, the suspension cell line acclimation method provided by the invention has the advantages that suspension acclimation culture is carried out while serum contents are lowered, cell passage number corresponding to each serum gradient can be maintained to be 1-3 times, acclimation of the adherent cells is controlled to be completed within one month on the whole, the acclimation period is shortened, and acclimation cost is saved. The suspension cell line obtained by adopting the suspension cell line acclimation method has good dispersibility, high viability and high growth rate.

Description

technical field [0001] The invention belongs to the technical field of cell culture, in particular to a suspension cell strain and a domestication method thereof. Background technique [0002] Animal cell culture, especially large-scale animal cell culture technology has been widely used in the production of biological products including monoclonal antibodies, vaccines, recombinant proteins and genetically engineered drugs. Due to the complex structure and high degree of post-translational modification of a large number of biological drugs, eukaryotic expression systems, especially animal cell expression systems, have become the main expression system for industrial production of biological drugs due to their high level of glycosylation and the degree of post-translational modification closest to humans. Since most animal cells have the characteristics of adherent culture, in the process of large-scale industrial production, the widely used bioreactor cascade amplification p...

AI Technical Summary

Problems solved by technology

However, often when the serum content is reduced from 5% to 0% in the square bottle, the phenomenon that the cell growth rate slows down gradually occurs, resulting in slow progress of the domestication process, and the domestication cycle is generally more than 6 months

It often takes more than 5 generations for the cells to adapt to each gradient of serum; especially when the serum content drops to 1% to 0%, the cell adhesion performance decreases, the metabolism is slow, and it is very easy for cells to die in clusters, resulting in the failure of domestication

Images