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Application of glycoside dipeptide conditioned medium in cryopreservation of mesenchymal stem cells and preparation of commercial cryopreservation agents

A technology of conditioned medium and glycerodipeptide, applied in the field of stem cells, can solve problems such as cell damage, and achieve the effect of improving the recovery rate of cryopreservation

Active Publication Date: 2021-10-01
广东创晟控股集团有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, in order to improve the cryopreservation and recovery rate of human amniotic fluid-derived mesenchymal stem cells, liquid nitrogen cryopreservation is basically used. However, there are many steps in the cryopreservation and recovery process of liquid nitrogen cryopreservation. Damage to cells cannot be ignored

Method used

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  • Application of glycoside dipeptide conditioned medium in cryopreservation of mesenchymal stem cells and preparation of commercial cryopreservation agents
  • Application of glycoside dipeptide conditioned medium in cryopreservation of mesenchymal stem cells and preparation of commercial cryopreservation agents
  • Application of glycoside dipeptide conditioned medium in cryopreservation of mesenchymal stem cells and preparation of commercial cryopreservation agents

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Cell cryopreservation, recovery and characteristic detection

[0026] 1. Experimental materials

[0027] Amniotic fluid mesenchymal stem cells HuFACs-11 were cryopreserved in liquid nitrogen in the laboratory and were originally purchased from Shanghai Yaji Biotech.

[0028] Calf serum was purchased from Shanghai Suer Biotechnology Co., Ltd., branded as Gibco.

[0029] Basic fibroblast growth factor was purchased from Sigma, USA.

[0030] Low-sugar DMEM medium L-DMEM was purchased from Gibco, USA.

[0031] FITC or PE-labeled mouse anti-human antibodies CD29, CD34, CD44, CD45, CD105, HLA-DR and isotype controls were purchased from BD Biosciences.

[0032] Glycerin dipeptide (CAS No. 658-79-7) was purchased from Wuhan Senditai Technology Co., Ltd., with a purity of 99%.

[0033] The brand of the programmed cooling box is Nalgene.

[0034] 2. Experimental method

[0035] 1. Culture of human amniotic fluid mesenchymal stem cells

[0036] Human amniotic flu...

Embodiment 2

[0073] Embodiment 2: A kind of glycoside dipeptide conditioned medium

[0074] A kind of glycoside dipeptide conditioned medium, which is composed of low-sugar DMEM medium and glycoside dipeptide, and the concentration of glycoside dipeptide is 100nM. The glycoside dipeptide conditioned medium can be used to prepare a cryoprotectant for human amniotic fluid mesenchymal stem cells.

Embodiment 3

[0075] Example 3: A cryoprotectant for human amniotic fluid mesenchymal stem cells

[0076] A human amniotic fluid mesenchymal stem cell cryopreservation agent is composed of low-sugar DMEM conditioned medium, calf serum and DMSO with a volume ratio of 5:4:1, and the low-sugar DMEM conditioned medium is low-sugar DMEM containing 100nM glycoside dipeptide Medium.

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Abstract

The invention discloses the application of the glycoside dipeptide conditioned medium in the cryopreservation of mesenchymal stem cells and the preparation of a commercial cryopreservation protective agent. The research of the present invention found that glycoside dipeptide can improve the cryopreservation recovery rate of human amniotic fluid mesenchymal stem cells without significantly affecting the phenotype, proliferation activity and multidirectional differentiation ability of human amniotic fluid mesenchymal stem cells. And the conditioned medium containing glycoside dipeptide can be used to prepare cryopreservation agent for human amniotic fluid mesenchymal stem cells.

Description

technical field [0001] The invention belongs to the field of stem cells and relates to cryopreservation of stem cells, in particular to the application of a glycoside dipeptide conditional medium in the cryopreservation of mesenchymal stem cells and the preparation of a commercial cryopreservation protective agent. Background technique [0002] Mesenchymal stem cells (MSCs) have multi-lineage differentiation potential and are ideal seed cells for cell therapy and gene therapy. Existing studies have shown that MSCs can be cultured in both bone marrow and umbilical cord blood, but bone marrow-derived MSCs have the possibility of virus contamination and the disadvantages of obvious decline in cell number, expansion and differentiation ability with age, while umbilical cord blood MSCs Cultivation has disadvantages such as low yield and instability, and its application is subject to certain restrictions. Recent studies have shown that amniotic fluid is a new source of MSCs. Comp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 马春华刘燕陆扬
Owner 广东创晟控股集团有限公司
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