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Mesenchymal stem cell frozen stock solution and preparation method thereof

A technique of mesenchymal stem cells and cryopreservation solution, applied in the field of mesenchymal stem cell cryopreservation solution and its preparation, can solve the problems of animal pathogen pollution, increase of operation steps, uncertain factors, cell damage, etc. Cell differentiation does not change, good stability

Active Publication Date: 2012-06-27
CHENGDU QINGKE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Toxic dimethyl sulfoxide (DMSO) and animal serum are mainly used in the cryopreservation of mesenchymal stem cells. DMSO is toxic and will inevitably cause certain damage to cells, while animal serum is contaminated by animal pathogens. Possibilities, which add operational steps and uncertainties to cell production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021]

[0022] Embodiment 2: Comparison of the effect of the cryopreservation solution of the present invention with that of the DMSO-containing cryopreservation solution

Embodiment 2

[0023] Freezing solution 1: Trehalose 200mM;

[0024] ATP 3mM;

[0025] Human Serum Albumin 0.025%;

[0026] According to the above concentrations, the above three substances were added to the mixed sugar electrolyte solution to prepare cryopreservation solution 1.

[0027] Freezing solution 2: DMSO volume ratio 10%;

[0028] Fetal bovine serum volume ratio 10%;

[0029] DMEM medium volume ratio 80%;

[0030] Prepare mixed freezing solution 2 according to the above ratio.

[0031] Steps:

[0032] 1) Prepare cryopreservation solution 1 with trehalose, ATP, human serum albumin, and mixed sugar electrolyte solution according to the above ratio; prepare cryopreservation solution 2 with DMSO, fetal bovine serum, and DMEM medium according to the above ratio;

[0033] 2) Use cryopreservation solution 1 and cryopreservation solution 2 to freeze adipose-derived mesenchymal stem cells, umbilical cord mesenchymal stem cells, and dental pulp mesenchymal stem cells, and put them...

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Abstract

The invention discloses a mesenchymal stem cell frozen stock solution which does not contain animal proteins and DMSO (dimethyl sulfoxide). The frozen stock solution comprises trehaloses, a carbohydrate and electrolyte injection, ATP (adenosine triphosphate) and human serum albumins. The frozen stock solution has the advantages of high recovery rate (90%), no influence on characteristics of stem cells, unchanged cell proliferation, unchanged cell differentiation, easy use, good stability and the like. The invention also discloses a preparation method of the frozen stock solution.

Description

technical field [0001] The application relates to a mesenchymal stem cell cryopreservation solution and a preparation method thereof, belonging to the field of biomedicine. Background technique [0002] Mesenchymal stem cells (mesenchymal stem cells, MSC) are important members of the stem cell family, derived from the mesoderm and ectoderm in the early stages of development. Originally found in bone marrow, MSCs 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 used as an ideal seed ce...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 郑榆坤海泉
Owner CHENGDU QINGKE BIOTECH
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