A kind of serum-free medium for cultivating adherent cells and application thereof
A technology of serum-free medium and adherent cells, which is applied in the direction of cell culture medium, culture process, animal cells, etc. It can solve the problems of death, failure to proliferate normally, and failure to use tumor cells. Effect
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[0058] The present invention has no special limitation on the preparation method of the DMEM high-glucose medium, and the well-known medium preparation scheme in the art can be adopted.
[0059] The serum-free medium provided by the invention is based on DMEM high-glucose medium and reduced-serum medium. The volume ratio of the DMEM high glucose medium and the reduced serum medium is 50-54:46-50, preferably 52:48. The present invention has no special limitation on the type of the serum-reduced medium, and the serum-reduced medium well known in the art can be used. In the embodiment of the present invention, the reduced serum medium is preferably Gibco Opti-MEM.
[0060] The serum-free medium provided by the present invention includes a serum replacement. Each liter of basal medium contains 1.5-2.5 ml of serum replacement, more preferably 2 ml. In the present invention, the serum substitute is preferably Gibco B-27. The serum substitute replaces serum components.
[0061] ...
Embodiment 1
[0067] Serum-free medium configuration. The serum-free medium is composed of the following components: DMEM high-glucose medium and reduced-serum medium as basal medium, each liter of basal medium contains 2ml of serum replacement, 76.8mg of complex amino acids, 1×10 5 U penicillin and 1 x 10 5 mg of streptomycin.
[0068] The volume ratio of DMEM high glucose medium and reduced serum medium was 52:48.
[0069] The formula of DMEM high glucose medium is as follows:
[0070]
[0071] Reduced serum medium Opti-MEM was purchased from Gibco.
[0072] Serum substitutes were purchased from gibco
[0073] The formula of compound amino acid is as follows:
[0074] components parts by weight Glycine 7.50 L-alanine 8.90 L-Asparagine 13.20 L-Aspartic Acid 13.30 L-glutamic acid 14.70 L-proline 11.50 L-serine 10.50
[0075] The serum-free medium was prepared as follows:
[0076]In a sterile environment, mix DMEM high-gluco...
Embodiment 2
[0078] Similar to the scheme of Example 1, the difference is that each liter of basal medium contains a volume concentration of 1.5ml serum replacement, 95.5mg compound amino acids, 1.1×10 5 U penicillin and streptomycin 110mg. The volume ratio of DMEM high glucose medium and reduced serum medium was 50:50. The serum-free medium was prepared according to the preparation method of Example 1.
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