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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

Active Publication Date: 2020-12-25
WUXI PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

And CN102352343A and CN104830765A disclosed culture medium scheme and fetal bovine serum substitute all are used for lymphocyte culture, and lymphocyte adopts suspension culture method, can't be used for the tumor cell of adherent culture
Therefore, in the above medium scheme, the tumor cells cannot adhere to the wall normally, nor can they proliferate normally. After a certain period of culture, the apoptosis rate increases significantly until all of them die, which affects the normal progress of experimental research.

Method used

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  • A kind of serum-free medium for cultivating adherent cells and application thereof
  • A kind of serum-free medium for cultivating adherent cells and application thereof
  • A kind of serum-free medium for cultivating adherent cells and application thereof

Examples

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preparation example Construction

[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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Abstract

The invention provides a serum-free culture medium for adherent cell culture and application of the serum-free culture medium, and belongs to the field of cell culture media. High-glucose DMEM (dulbecco's modified eagle media) and serum-reduced culture media are used as basic culture media, and the basic culture media contain serum substitutes, compound amino acid, penicillin and streptomycin. Theserum-free culture medium and the application have the advantages that the serum-free culture medium is free of cell culture media with animal-derived substances and accordingly is independent from animal serum; the serum-free culture medium comprises definite components, an alternative scheme for serum culture media is provided, and accordingly the serum-free culture medium is favorable for market promotion; the serum-free culture medium can be applied to the adherent cell culture; self-renewal of tumor cells and normal cell lines which are subjected to adherent culture can be maintained bythe serum-free culture medium, and long-time culture and passage can be implemented without increase of cell apoptosis proportions.

Description

technical field [0001] The invention belongs to the field of cell culture medium, and in particular relates to a serum-free culture medium for culturing adherent cells and an application thereof. Background technique [0002] Cell culture is the most commonly used basic technique in cell biology and tumor-related research. When cells are cultured in vitro, the medium provides the necessary external environment for cell growth, division, and proliferation, including buffers that simulate physiological acid-base concentrations, substances required for basic metabolism, and some nutritional factors. According to cell characteristics, cell culture methods are usually divided into suspension culture and adherent culture. Except for stem cells, lymphocytes and other blood system cells commonly used in suspension culture, most cells in the body, including normal cells and tumor cells, rely on adherence. wall culture. When culturing adherent cells, in addition to providing the cul...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/00C12N5/09
CPCC12N5/0037C12N5/0693C12N2500/32C12N2500/84
Inventor 邹健纪丽邹奕苏殷莹汪京京龚玲丽胡亚玲
Owner WUXI PEOPLES HOSPITAL
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