Cryopreservation solution and cryopreservation method for human amniotic mesenchymal stem cells

A kind of stem cell and human amniotic membrane technology, applied in the field of cell culture, can solve the problem of low cell survival rate, achieve high cell survival rate, ensure cell activity, and improve the effect of survival rate

Active Publication Date: 2020-06-09
瑞因细胞工程科技(广州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current serum-free cryopreservation medium for human amniotic mesenchymal stem cells has a low survival rate of recovered cells, which cannot meet the needs of clinical application.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A cryopreservation solution for human amniotic mesenchymal stem cells, which consists of the following raw materials: DMEM / F12 medium, dimethyl sulfoxide, pullulan, retinol-binding protein, Ascophyllum nodosum extract, γ-oryzanol, Tea polyphenols.

[0026] Wherein the volume ratio of DMEM / F12 medium and dimethyl sulfoxide is 100: 1.2, the ratio of pullulan, retinol-binding protein, Ascophyllum nodosum extract, γ-oryzanol, tea polyphenols in the cryopreservation solution The concentrations were: pullulan 2.7mg / mL, retinol binding protein 8.5μg / mL, Ascophyllum nodosum extract 2.0mg / mL, γ-oryzanol 10.5μg / mL, tea polyphenol 17.0μg / mL.

[0027] A cryopreservation method for human amniotic mesenchymal stem cells, comprising the following steps: adding dimethyl sulfoxide, Ascophyllum nodosum extract, γ-oryzanol, and tea polyphenols to DMEM / F12 medium, adding P3 generation human amniotic membrane Mesenchymal stem cells, add 1.0×10 cells per ml of culture medium 7 Pre-cool at ...

Embodiment 2

[0029] A cryopreservation solution for human amniotic mesenchymal stem cells, which consists of the following raw materials: DMEM / F12 medium, dimethyl sulfoxide, pullulan, retinol-binding protein, Ascophyllum nodosum extract, γ-oryzanol, Tea polyphenols.

[0030] Wherein the volume ratio of DMEM / F12 medium and dimethyl sulfoxide is 100:1, and the volume ratio of pullulan, retinol-binding protein, Ascophyllum nodosum extract, γ-oryzanol, and tea polyphenols in the cryopreservation solution The concentrations were: pullulan 2.5 mg / mL, retinol binding protein 8.0 μg / mL, Ascophyllum nodosum extract 1.5 mg / mL, γ-oryzanol 10.0 μg / mL, tea polyphenol 15.0 μg / mL.

[0031] A cryopreservation method for human amniotic mesenchymal stem cells, comprising the following steps: adding dimethyl sulfoxide, Ascophyllum nodosum extract, γ-oryzanol, and tea polyphenols to DMEM / F12 medium, adding P4 generation human amniotic membrane For mesenchymal stem cells, add 1.2×10 cells per ml of culture m...

Embodiment 3

[0033] A cryopreservation solution for human amniotic mesenchymal stem cells, which consists of the following raw materials: DMEM / F12 medium, dimethyl sulfoxide, pullulan, retinol-binding protein, Ascophyllum nodosum extract, γ-oryzanol, Tea polyphenols.

[0034] Wherein the volume ratio of DMEM / F12 medium and dimethyl sulfoxide is 100:1.5, and the ratio of pullulan, retinol-binding protein, Ascophyllum nodosum extract, γ-oryzanol, and tea polyphenols in the cryopreservation solution The concentrations were: pullulan 2.5 mg / mL, retinol binding protein 8.7 μg / mL, Ascophyllum nodosum extract 2.3 mg / mL, γ-oryzanol 12.0 μg / mL, tea polyphenol 18.0 μg / mL.

[0035] A cryopreservation method for human amniotic mesenchymal stem cells, comprising the following steps: adding dimethyl sulfoxide, Ascophyllum nodosum extract, γ-oryzanol, and tea polyphenols to DMEM / F12 medium, adding P5 generation human amniotic membrane For mesenchymal stem cells, add 1.5×10 cells per ml of culture medium...

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PUM

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Abstract

The invention discloses a cryopreservation solution for human amniotic mesenchymal stem cells. The cryopreservation solution is prepared from the following raw materials: a basal culture medium, dimethyl sulfoxide, pullulan, retinol binding protein, an ascophyllum nodosum extract, gamma-oryzanol and tea polyphenol. The cryopreservation solution for the human amniotic mesenchymal stem cells provided by the invention has the following advantages: the culture medium is used for providing nutrition for cells; the pulullan, the retinol binding protein, the ascophyllum nodosum extract, the gamma-oryzanol and the tea polyphenol are also added into the cryopreservation solution; the above-mentioned components have a synergistic effect in the cell cryopreservation process; the addition of exogenousserum is not needed; the use amount of dimethyl sulfoxide is reduced; meanwhile, the activity of cells after cryopreservation is ensured; the cells are rapidly recovered; and the survival rate of recovered cells is high. The invention also provides a cryopreservation method for the human amniotic mesenchymal stem cells. The components of the cryopreservation solution are added in batches according to temperature changes in the cryopreservation process; damage of the ice crystals inside and outside the cells to the cells is further reduced; the survival rate of the recovered cells is increased; and the cryopreservation method is simple, convenient and easy to operate.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a cryopreservation liquid for human amniotic mesenchymal stem cells and a cryopreservation method thereof. Background technique [0002] Mesenchymal stem cells (Mesenchymal Stem Cells, MSCs) are a kind of pluripotent stem cells, which have all the common characteristics of stem cells, that is, self-replication ability and multi-lineage differentiation ability. Under specific induction conditions in vivo or in vitro, they can differentiate into neural, Vascular endothelium, cartilage, muscle, liver, cardiac muscle and other tissue cells. MSCs have become the seed cells for cell and gene therapy research because of their relative lack of immunogenicity, simple culture technology, and cryopreservation. They have potential clinical application value and have become a hot spot in stem cell research. [0003] Cell cryopreservation will significantly change the thermodynamic, chem...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 李刚霍亚如黄金刚
Owner 瑞因细胞工程科技(广州)有限公司
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