Preparation method of adipose tissue-derived stem cell secreted factor

Abstract

The invention discloses a preparation method of adipose-derived mesenchymal stem cell secreted factors, and relates to the technical field of mesenchymal stem cell secreted factor extraction, the preparation method comprises the following steps: 1, cell recovery; step 2, subculture: carrying out subculture on the adipose-derived stem cells by using a subculture medium; step 3, harvesting secretory factors; the subculture medium in the step 2 is prepared from 400 to 600 ml / L of stem cell basal culture medium, 1 to 3 mmol / L of L-alanyl-L-glutamine, 10 to 20 [mu] mol / L of 4-2-hydroxyethyl-1-piperazine ethanesulfonic acid, 2.0 to 6.0 [mu] g / mL of insulin, 10 to 20 g / L of D-glucose, 10 to 100 [mu] mol / L of L-ascorbic acid, 30 to 50 [mu] g / mL of all-transretinoic acid, 0.3 to 0.8 [mu] g / mL of serum protein, 0.1 to 0.3 [mu] g / mL of tryptase and 10.0 to 20.0 [mu] g / mL of ginsenoside. The adipose-derived mesenchymal stem cell secreted factor obtained in the application process has good stability, is not easily influenced by allergens, and can exert a good and stable tissue repair effect.

Description

technical field [0001] The application relates to the field of extracting secreted factors of mesenchymal stem cells, more specifically, it relates to a preparation method of secreted factors of adipose-derived mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (mesenchymal stem cells, MSCs) are a type of pluripotent stem cells belonging to the mesoderm, which have strong proliferation ability and multi-lineage differentiation potential. Adipocytes also have the ability to differentiate into muscle cells, liver cells, hematopoietic cells, nerve cells, pancreatic islet cells, and other cells. MS cells can differentiate into specific cells in a suitable environment to repair damaged tissues, and paracrine effects in Played a greater role in the treatment of tissue damage, that is, MSC secreted cytokines played a large role in tissue repair. [0003] In related technologies, during the extraction process of mesenchymal stem cell secreted factors, thir...

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Problems solved by technology

[0004] In view of the above-mentioned related technologies, the applicant believes that the addition of human serum albumin can make the mesenchymal stem cell (MSC) secreted factors in the supernatant more stable, but the mesenchymal stem cell secreted factors are mostly used clinically for allergic reactions. For the treatment of diseases, when MSC secreted factors are applied to the user's affected area and used for tissue repair, they have been affected by allergens during use, resulting in a greatly reduced overall stability, which in turn leads to poor tissue repair effects. Therefore, A new solution needs to be proposed to solve the above problems

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