Serum-free culture medium for adherent cell culture and application of serum-free culture medium
A technology of serum-free culture medium and adherent cells, applied in cell culture medium, culture process, tissue culture, etc., can solve the problems of death, failure to proliferate normally, failure to use tumor cells, etc., to achieve component determination and facilitate market promotion Effect
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[0057] 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.
[0058] 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.
[0059] 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.
[0060] ...
Embodiment 1
[0066] 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.
[0067] The volume ratio of DMEM high glucose medium and reduced serum medium was 52:48.
[0068] The formula of DMEM high glucose medium is as follows:
[0069]
[0070] Reduced serum medium Opti-MEM was purchased from Gibco.
[0071] Serum substitutes were purchased from gibco
[0072] The formula of compound amino acid is as follows:
[0073] components
parts by weight
7.50
L-alanine
8.90
13.20
13.30
14.70
L-proline
11.50
L-serine
10.50
[0074] The serum-free medium was prepared as follows:
[0075]In a ster...
Embodiment 2
[0077] 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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