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Clinic-level adipose-derived stem cell preparation and storage methods

A technology of adipose stem cells and storage methods, which is applied in the field of preparation and storage of clinical-grade adipose stem cells, which can solve the problems of low yield rate of adipose tissue, ADSCs animal-derived protein pollution, and easy aging, so as to reduce the link of in vitro culture, Maintain cell phenotype and doubling time, plateau prolongation effect

Inactive Publication Date: 2015-12-09
丛秀丽
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Problems solved by technology

[0022] The technical problem to be solved by the present invention is to overcome the defects in the prior art that the separation and cultivation of ADSCs are easily contaminated by animal-derived proteins, the output rate of adipose tissue is low, and it is easy to age, and provide a preparation and preparation of clinical-grade adipose stem cells. storage method

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  • Clinic-level adipose-derived stem cell preparation and storage methods
  • Clinic-level adipose-derived stem cell preparation and storage methods
  • Clinic-level adipose-derived stem cell preparation and storage methods

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Embodiment Construction

[0053] Preferred embodiments of the present invention are described below, and it should be understood that the preferred embodiments described here are only used to illustrate and explain the present invention, and are not intended to limit the present invention.

[0054] A method for preparing clinical grade adipose stem cells, comprising the following steps:

[0055] (1), the preparation of the platelet lysate PL of pathogen inactivation

[0056] Collect autologous platelets by machine or collect expired platelets from the central station, centrifuge at 20-25°C, 1200-1500rpm for 10-15min to inactivate pathogens;

[0057] Perform 3-5 consecutive cycles of -80°C freezing and 37°C recovery to fully lyse the platelets, centrifuge at 4000rpm for 30 minutes, collect the platelet lysate, freeze at -80°C for later use, and conduct pathogenic testing during the freezing period;

[0058] The content of cytokines in PL was measured by ELISA method, and the results were as follows: f...

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Abstract

The invention discloses clinic-level adipose-derived stem cell preparation and storage methods. Mesenchymal stem cell culture of a fat tissue source is performed by adopting a serum-free complete medium without animal source protein pollution, safety and effectiveness are realized, immunogenicity is avoided, the rate of increase of ADSCs can be better maintained, a plateau period is prolonged, an expansion multiple of each generation of cells is greatly improved, and the clinic treatment number is ensured; the serum-free complete medium is applied to preparation of clinic-level ADSCs for the first time; a fat tissue is collected from the abdomen and can be taken discontinuously repeatedly, thus avoiding damage to the adipose-derived stem cells in a liposuction process; the ADSCs are prepared by using a wall adherence method, thus increasing the primary cell yield and lowering the cost; an adopted novel serum-free cell frozen preservation solution meets a clinical application standard, reduces the use amount of DMSO, does not obviously influence the motility rate, phenotype and doubling time of the ADSCs, can be directly used in clinic infusion after reviving, is convenient to transport, and ensures the safety of clinic treatment.

Description

technical field [0001] The invention relates to the application field of stem cells, in particular to a method for preparing and storing clinical-grade fat stem cells. Background technique [0002] Mesenchymal stem cells: are important members of the stem cell family, derived from the mesoderm in the early development stage, and belong to pluripotent stem cells. Mesenchymal stem cells were originally discovered in the bone marrow, and have attracted increasing attention because of their multi-lineage differentiation potential, hematopoietic support and promotion of stem cell implantation, immune regulation and self-replication. For example, mesenchymal stem cells can be differentiated into fat, bone, cartilage, muscle, tendon, ligament, nerve, liver, cardiac muscle, endothelial and other tissue cells under specific induction conditions in vivo or in vitro, after continuous subculture and cryopreservation It still has multi-directional differentiation potential and can be us...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A01N1/02
Inventor 丛秀丽
Owner 丛秀丽
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