Suspension acclimatization and serum-free acclimatization method for HEK (human embryonic kidney)-293T cells

Abstract

The invention relates to a suspension acclimatization and serum-free acclimatization method for HEK (human embryonic kidney)-293T cells, and belongs to the field of biotechnology. The suspension acclimatization and serum-free acclimatization method includes acclimatizing the HEK-293T cells adaptive to adherent culture so that the HEK-293T cells are adaptive to suspension culture in culture media with serum; and acclimatizing the HEK-293T cells adaptive to suspension culture in the culture media with the serum in a serum reducing mode so that the HEK-293T cells are adaptive to serum-free acclimatization. The suspension acclimatization and serum-free acclimatization method has the advantages that the growth status of the HEK-293T cells cultured by the suspension acclimatization and serum-free acclimatization method is stable, the HEK-293T cells are high in dispersibility and are completely adaptive to suspension culture in the serum-free culture media with certain chemical compositions, and batch production can be carried out.

Description

【Technical field】 [0001] The invention relates to a method for culturing HEK-293T cells, in particular to a method for suspension and serum-free domestication of HEK-293T cells, and belongs to the field of biotechnology. 【Background technique】 [0002] Mammalian cell large-scale culture technology is one of the downstream common technologies of biopharmaceutical companies, and it plays an extremely important role in this industry. The key to this technology is to achieve suspension and serum-free culture of mammalian cells, thereby increasing production capacity and reducing costs. Because the usual practice in small-scale cell culture is to add serum to the medium, and the chemical composition of serum is uncertain, and the quality is unstable, and there are batch-to-batch differences. The quality of biological products produced with such cells is difficult to control and difficult to pass. Compared with the common adherent culture, suspension culture can grow more cells p...

AI Technical Summary

Problems solved by technology

The disadvantages are: 1. The quality of the serum is unstable, there is a difference between batches, and the chemical composition of the serum is uncertain. If the biological products produced by such cells are difficult to control, it is difficult to pass the approval of the Food and Drug Administration; 2. .Due to its adherent nature, compared with suspension culture cells, the number of cells that can grow per unit volume is small, so it is difficult to achieve large-scale production

[0006] However, the existing suspension domestication technology has the following defects: 1. Some cannot achieve complete serum-free culture; 2. Generally, a two-step method is adopted, first serum weaning and then suspension culture

Since the medium used for serum weaning is usually basal medium plus serum, the nutrient content of the basal medium is weak, and the surface area / volume ratio of the adherent culture during the acclimatization process is usually small, which limits the intake of nutrients, and the entire acclimatization process takes a long time