High-density continous pouring culture of animal cell

Abstract

The invention opened a high density filling culture method of the animal cells. It study the metabolic rule of the cell growth, the nutrient consumption and the metabolic byproduct accumulation by studying theirs dynamics to determine and control the optimal nutritive condition. It feed the nutrient quality of cell growth by the stoichiometric relationship and the filling culture. The process can control the nutrient in a proper low concentration, so we can get the high density of the cells and the product concentration.

Description

technical field [0001] The invention relates to a large-scale, high-density, serum-free continuous perfusion culture method for animal cells. technical background [0002] The cultivation process of animal cells in the bioreactor can have different operation modes, such as batch culture (Batch), continuous culture (Continuous), perfusion culture (Perfusion) and fed-batch culture (Fed-batch), etc., different operation modes has its different characteristics. [0003] Continuous perfusion culture is to add fresh nutrient solution at a certain rate during the culture process, and discharge the supernatant at the same rate. The fragments are separated, and the living cells are returned to the reactor to continue culturing. Continuous perfusion culture not only maintains a relatively constant culture state, avoids the accumulation of harmful metabolic by-products, but also enables the cells in the reactor to reach a relatively high density. [0004] In recent years, animal cel...

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Problems solved by technology

However, there are still some deficiencies in the research work of these scholars. The main reason is that they often only consider glucose and / or glutamine. Limiting factors, in the end, often cause a large number of cells to die, the number of dead cells is greater than the number of living cells, and the increase in the density of living cells and product concentration is still limited

Although the medium continuous perfusion culture has more advantages than fed-batch culture, in addition to continuously supplementing nutrients, it can also remove metabolic byproducts in time, so the obtained cell density can be increased by one to two orders of magnitude compared with batch culture, but the current Perfusion culture for research and application, because its medium is unbalanced, often at the expense of high-speed medium perfusion, and the product concentration is diluted in large quantities, which is not worth the economic gain

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