Expi293F cell culture method

Abstract

The invention discloses an Expi293F cell culture method, which comprises the following steps: inoculating Expi293F cells into a bioreactor for culture, and carrying out perfusion dynamic culture for 32-48 hours: uniformly dividing a culture solution to be perfused into a plurality of strands, and perfusing into the bioreactor along the inner wall of the bioreactor in a wall-attached manner, with the stirring speed in the bioreactor being 80-90 rpm and the perfusion rate of the perfused solution being 0.5-2 working volumes / d, during dynamic perfusion culture, the culture conditions in the reactor are kept as follows: the temperature is 36.5-37 DEG C, the DO is 40-60%, and the pH is 7.2 + / -0.2; a serum-free culture solution used in the culture process is prepared from 95wt% of 24.75 g / L Dynami culture medium, 4wt% of 200mM glutamine and 1wt% of an anti-caking agent. According to the invention, the perfusion mode is improved, and proper perfusion rate and stirring strength reduction are matched, so that the injection of the culture medium in the perfusion process also plays roles in expanding mass transfer, reducing the influence of cell agglomeration on cell metabolism and reducing physical damage to cells, high-density cells and high survival rate are obtained, and remarkable effects are achieved.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to an Expi293F cell culture method. Background technique [0002] Cell culture refers to a method that simulates the in vivo environment (sterility, suitable temperature, pH and certain nutrient conditions, etc.) in vitro to make it survive, grow, reproduce and maintain its main structure and function. For large-scale cell culture, bioreactors are often used. A bioreactor is a container that uses microorganisms, plants, animals or human cells to convert raw materials into products through biological methods. [0003] The cell culture bioreactor can carry out large-scale cell culture. Its advantages are that it can realize automatic control and real-time monitoring of culture conditions during the cell culture process, and can relatively stably adjust the entire production conditions, thereby improving the stability of product quality. sex. However, in the actual production process, wh...

AI Technical Summary

Problems solved by technology

However, in the actual production process, when the volume of the reactor increases, in order to maintain the uniformity of the environment, it is necessary to increase the intensity of the stirring, but this brings higher shear rate and shear force and causes the cells to be physically damaged; but if Therefore, reducing the stirring intensity is not conducive to mass transfer

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