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Mesenchymalstem cellfrozen stock solution as well as preparation method and using method thereof

A technique for mesenchymal stem cells and cryopreservation solution, which is applied in the field of mesenchymal stem cell cryopreservation solution and its preparation, can solve the problems of unsatisfactory long-term cryopreservation effect and complex components, and achieves the effect of good cryopreservation effect and clear composition.

Inactive Publication Date: 2018-12-28
深圳市浊安认证生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0021] In order to solve the problems existing in the prior art, the present invention provides a mesenchymal stem cell Cryopreservation solution and its preparation method and use method. The mesenchymal stem cell cryopreservation solution of the present invention is relatively simple, suitable for popularization and application, has low toxicity, and can store mesenchymal stem cells for a long time while maintaining its biological characteristics.

Method used

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  • Mesenchymalstem cellfrozen stock solution as well as preparation method and using method thereof
  • Mesenchymalstem cellfrozen stock solution as well as preparation method and using method thereof
  • Mesenchymalstem cellfrozen stock solution as well as preparation method and using method thereof

Examples

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Effect test

Embodiment 1

[0055] This example provides a mesenchymal stem cell cryopreservation solution, including 5% of dimethyl sulfoxide, 10% of human albumin, 75% of DMEM / F-12 medium, and 5umol of Rock inhibitor Y27632 / L and hydroxyethyl starch is 6%. Dimethyl sulfoxide is 5%, human albumin is 10%, DMEM / F-12 medium is 75%, Rock inhibitor Y27632 is 5umol / L and hydroxyethyl starch is 6%.

[0056] The preparation method is as follows: according to the cell density and the volume of the freezing solution to be prepared, first take the corresponding proportion of hydroxyethyl starch, Rock inhibitor Y27632 and serum-free stem cell medium, and gently suspend; then add the corresponding proportion of human serum albumin Suspend the protein gently; finally add dimethyl sulfoxide drop by drop and mix well to obtain the mesenchymal stem cell cryopreservation solution.

[0057]The method of use is: collect the cells to be frozen, digest, centrifuge, and discard the supernatant; then add sterile PBS to the t...

Embodiment 2

[0059] This example provides a mesenchymal stem cell cryopreservation solution, including 5% dimethyl sulfoxide, 10% human serum albumin, 70% DMEM / F-12 medium, and 5umol of Rock inhibitor Y27632 / L and hydroxyethyl starch is 6%.

[0060] The preparation method is as follows: according to the cell density and the volume of the frozen storage solution to be prepared, first take the corresponding proportion of hydroxyethyl starch, Rock inhibitor Y27632 and serum-free stem cell medium, and gently suspend; then add the corresponding proportion of human serum albumin Suspend protein gently; finally add dimethyl sulfoxide drop by drop, mix well, put dimethyl sulfoxide in 4°C refrigerator for 10-30 minutes before dropping, and pre-cool to 4°C, that is It is the cryopreservation liquid for the mesenchymal stem cells.

[0061] The method of use is: collect the cells to be frozen, digest, centrifuge, and discard the supernatant; then add sterile PBS to the treated cells to be frozen, co...

experiment example 1

[0063] Study the effect of the mesenchymal stem cell cryopreservation solution of the present invention on the cryopreservation and recovery of human umbilical cord mesenchymal stem cells

[0064] 1. Experimental purpose: compare and evaluate the effect of the mesenchymal stem cell cryopreservation solution of the present invention and the common cryopreservation solution on the cryopreservation and recovery of human umbilical cord mesenchymal stem cells

[0065] 2. Experimental method: In this experimental method, UC-MSCs were cryopreserved for 1, 3, and 6 months by using different cryopreservation solutions, and then recovered by trypan blue counting, flow cytometry phenotype detection, and CCK8 analysis of cell viability. Proliferation ability, to evaluate the cryopreservation effect of these two cell cryopreservation solutions on UC-MSCs.

[0066] The specific experimental operations involved are as follows:

[0067] (1) Cell culture method: Inoculate the collected UC-MSC...

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Abstract

The invention relates to a mesenchymalstem cellfrozen stock solution as well as a preparation method and a using method thereof. The mesenchymalstem cellfrozen stock solution comprises a Rock inhibitor Y27632, dimethyl sulfoxide, human serum albumin, hydroxyethyl starch and the like. The mesenchymalstem cellfrozen stock solution disclosed by the invention is applicable to a slow cryopreservation method; not only is the mesenchymalstem cellfrozen stock solution simple to operate, but also the formula has the advantages of low toxicity and low economic cost; the activity of mesenchymalstem cellscan also be kept for long time; and the mesenchymalstem cellfrozen stock solution has an excellent cryopreservation effect and is safer and more reliable.

Description

technical field [0001] The invention belongs to the field of cell technology, and in particular relates to a mesenchymal stem cell cryopreservation solution and a preparation method and application method thereof. Background technique [0002] Mesenchymal stem cells (MSCs) are a group of adult stem cells with high self-renewal ability and differentiation potential. They have attracted much attention because of their advantages such as immune regulation, cytokine secretion, and convenient extraction. Ideal seed cells. According to the data of Clinical Trials, the current clinical trial database in the United States, clinical trials related to mesenchymal stem cells have involved more than 300 diseases, and more than 650 clinical trials related to mesenchymal stem cells have been carried out in the United States alone. The main diseases include bone injury, neurodegenerative disease, diabetes, myocardial ischemia, myasthenia gravis, liver injury, etc. It can be seen that the...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 魏志璋黃馨萍
Owner 深圳市浊安认证生物技术有限公司
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