Serum-free medium for full-suspension culture of MDCK cells (madin-darby canine kidney cells) and preparation method of serum-free medium

Abstract

The invention discloses a serum-free medium for full-suspension culture of MDCK cells (madin-darby canine kidney cells) and a preparation method of the serum-free medium. The serum-free medium for full-suspension culture of the MDCK cells consists of a basic metabolism nutrient, nucleotide, vitamins, inorganic salt, a shear force protective agent, a cell clustering resisting agent, a pH buffer agent, a pH indicator, an influenza virus proliferation accelerant and other additives; and the preparation method of the serum-free medium for full-suspension culture of the MDCK cells comprises the following steps: 1) preparing mixed liquid: dissolving and mixing the raw materials; and 2) regulating pH: regulating pH of the mixed liquid to 6.3-6.7, and setting a constant volume, so that the serum-free medium for full-suspension culture of the MDCK cells is obtained. The medium supports the high-density full-suspension culture of the MDCK single cells and is capable of greatly shortening a time of domesticating the MDCK cells from adherent cells into the serum-free full-suspension cells; and the medium is applicable to the large-scale production of biological products, in particular veterinary biological products.

Description

technical field [0001] The invention relates to the field of preparation of a serum-free medium, in particular to a serum-free medium for full suspension culture of MDCK cells and a preparation method thereof. Background technique [0002] Canine kidney epithelial cells (Madin-Darby canine kidney cells, MDCK) are regarded as one of the most suitable cell lines for the production of influenza A and B influenza virus vaccines, and can be used to replace chicken embryos to culture influenza viruses. At present, a microcarrier adherent culture production process has been developed that utilizes MDCK cell lines instead of chicken embryos to cultivate influenza virus. Although this process has a certain production scale that can be achieved, it still has certain defects: 1. Microcarriers are difficult to multiply 2. The culture density of adherent cells is limited by the adherent area, resulting in a decrease in virus production; 3. Serum is usually added to help cell attachment a...

AI Technical Summary

Problems solved by technology

At present, a microcarrier adherent culture production process has been developed that utilizes MDCK cell lines instead of chicken embryos to cultivate influenza virus. Although this process has a certain production scale that can be achieved, it still has certain defects: 1. Microcarriers are difficult to multiply 2. The culture density of adherent cells is limited by the adherent area, resulting in a decrease in virus production; 3. Serum is usually added to help cell attachment and growth in adherent culture, and medium replacement is required , the process is complicated, and it will introduce mycoplasma, chlamydia or animal protein pollution, which poses a potential threat to the safety of vaccine products

Among the existing technologies, the serum-free medium contains components such as transferrin and other relatively expensive animal-derived proteins, which is not conducive to the development of veterinary biological products

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