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Preservation method of mesenchymal stem cell

A technology of stromal stem cells and preservation method, applied in the field of mesenchymal stem cell preservation, can solve the problems of slow growth rate of stem cells, and achieve the effects of long-term transportation preservation, simple and convenient preservation, and fast growth rate.

Inactive Publication Date: 2018-06-08
TIANJIN PURUI SAIER BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the existing technology can store mesenchymal stem cells at low temperature, the survival rate of stem cells during recovery is only about 80%, and the growth rate of stem cells after recovery is relatively slow

Method used

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  • Preservation method of mesenchymal stem cell
  • Preservation method of mesenchymal stem cell
  • Preservation method of mesenchymal stem cell

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0024] The preparation method of the cryopreservation solution is to dissolve dimethyl sulfoxide, sodium citrate, and citric acid in the solution medium, then add vitamin C injection to the solution medium, and then add human plasma and human blood albumin, and finally add low molecular weight heparin calcium to prepare the required cryopreservation solution;

[0025] Further, the pH value of the cryopreservation solution is 7.2-7.4.

[0026] Further, the solution medium is one or a combination of compound electrolyte injection or physiological saline.

[0027] Further, the mesenchymal stem cells are umbilical cord-derived mesenchymal stem cells.

[0028] Further, the concentration of the vitamin C is 50mg / ml.

[0029] Further, the concentration of the stem cell growth factor is 5-30 ng / mL

Embodiment 1

[0030] Example 1: The protective effect of vitamin C injection on mesenchymal stem cells

[0031] Group 1: Injection without vitamin C, human plasma, human serum albumin, low molecular weight heparin calcium, and the solution medium are uniformly mixed.

[0032] Group 2: only glucose, glucose, human plasma, human serum albumin, low molecular weight heparin calcium, and the solution medium are evenly mixed.

[0033] Put group 1 and group 2 into 2ml cryopreservation tubes respectively, and the number of mesenchymal stem cells is 3×10 6 / ml, prepare 6 tubes respectively, 3 tubes as a group, set up 6 groups and store at 4°C for 8h, 12h, 24h;

[0034] The above-mentioned mesenchymal stem cell sample solutions of each group were tested for viability, and the test results were as follows:

[0035]

[0036]

[0037] It can be seen from the above table that adding vitamin C injection can increase the survival rate of mesenchymal stem cells to a certain extent and play a protect...

Embodiment 2

[0038] Example 2: Effects of different concentrations of dimethyl sulfoxide on the growth rate of mesenchymal stem cells

[0039] Group 1: Dissolve 5% dimethyl sulfoxide, sodium citrate, and citric acid in the solution medium, then add vitamin C injection to the solution medium, then add human plasma and human serum albumin, and finally Add low-molecular-weight heparin calcium to prepare the required cryopreservation solution;

[0040] Group 2: Dissolve 10% dimethyl sulfoxide, sodium citrate, and citric acid in the solution medium, then add vitamin C injection to the solution medium, then add human plasma and human serum albumin, and finally Add low-molecular-weight heparin calcium to prepare the required cryopreservation solution;

[0041] Group 3: Dissolve 15% dimethyl sulfoxide, sodium citrate, and citric acid in the solution medium, then add vitamin C injection to the solution medium, then add human plasma and human serum albumin, and finally Add low-molecular-weight hep...

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Abstract

The invention discloses a preservation method of a mesenchymal stem cell. The method comprises freezing preservation liquid. A preparation method of the freezing preservation liquid comprises the steps of dissolving dimethyl sulfoxide, sodium citrate and citric acid in a solution medium, adding a vitamin C injection into the solution medium, adding human plasma and human serum albumin, and at last, adding low molecular weight calcium heparin to form the required freezing preservation liquid. The invention aims at providing the preservation method of the mesenchymal stem cell with a higher survival rate and a higher growth rate after thawing.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a preservation method of mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells are derived from the mesoderm in the early stage of development and belong to pluripotent stem cells. They were originally found in the bone marrow and have the advantages of multidirectional differentiation potential, hematopoietic support and promotion of stem cell implantation, immune regulation and self-replication. Low temperature or cryopreservation technology is a technology developed rapidly in the 1970s to preserve biological materials such as biological tissues, organs and cells under the condition of dry ice or liquid nitrogen. Under low temperature conditions, biological tissues are in a "dormant state", which ensures the genetic and morphological stability of biological materials, and is an effective method for preserving biological materials. In ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0215A01N1/0221A01N1/0226
Inventor 鲁振宇韩洪起张冰晶徐悦秦臻王瑶
Owner TIANJIN PURUI SAIER BIOLOGICAL TECH CO LTD
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