Cell freezing medium for human adipose-deprived stem cells

A technology of human adipose stem cells and cryopreservation solution, which is applied in the field of human adipose stem cell cryopreservation solution and its preparation, can solve the problems of cytotoxicity and stem cell damage, and achieve the effect of simple and feasible operation, reasonable price, and improved survival rate of resuscitation

Active Publication Date: 2016-04-20
北京九思九如健康科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, DMSO can be toxic to cells and cause irreversible damage to cryopreserved stem cells

Method used

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  • Cell freezing medium for human adipose-deprived stem cells
  • Cell freezing medium for human adipose-deprived stem cells
  • Cell freezing medium for human adipose-deprived stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] The preparation of embodiment 1 polypeptide

[0020] Take 5g of the leaves of Phoebe chinensis, clean them, mince them, squeeze the juice, add papain and trypsin, the amount of enzymes added is 8000IU / g leaves, the enzymolysis temperature is 47°C, the pH value is 7.5, the enzymolysis time is 1.5h, and the enzymolysis After completion, inactivate the enzyme at 90°C for 10 minutes; filter the insoluble matter after inactivating the enzyme to obtain a solution, add 4% activated carbon to the obtained polypeptide solution for adsorption and decolorization, and use dextran G-50 (SephadexG-50) for polypeptide separation, 20mmol / LHCl Solution elution, flow rate 1.3mL / min, collect eluted products in different time periods, adjust the solution to pH 7.0, centrifuge at 10,000 rpm for 15 minutes, desalt with macroporous resin DA201-C, concentrate in vacuo, and The supernatant was freeze-dried for later use; after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)...

Embodiment 2

[0021] Embodiment 2 Preparation of Human Adipose Stem Cell Cryopreservation Solution

[0022] 0.5% w / v LDL, 1% w / v trehalose, 1% glycerin, 1% w / v lecithin, 0.1% w / v polypeptide, bFGF1% w / v, vitamin E1% w / v, reduced glutathione 0.5% w / v was finally adjusted to 100 ml with DMEM medium, wherein the sequence of the polypeptide is shown in SEQ ID NO:1.

Embodiment 3

[0023] Example 3 Preparation of Human Adipose Stem Cell Cryopreservation Solution

[0024] 0.5% w / v LDL, 1% w / v trehalose, 1% glycerin, 1% w / v lecithin, 0.1% w / v polypeptide, bFGF1% w / v, vitamin E1% w / v, reduced glutathione 0.5% w / v was finally adjusted to 100 ml with DMEM medium, wherein the sequence of the polypeptide is shown in SEQ ID NO:2.

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Abstract

The invention provides a cell freezing medium for human adipose-deprived stem cells and a preparation method of the cell freezing medium. The cell freezing medium comprises low-density lipoprotein, trehalose, glycerin, lecithin, polypeptide, bFGF (fibroblast growth factor), vitamin E and reduced glutathione. The cell freezing medium is used for freezing the human adipose-deprived stem cells, especially can increase survival rate of frozen cells after recovery and has a good application prospect.

Description

technical field [0001] The invention relates to the technical field of tissue cell culture, in particular to a human adipose stem cell cryopreservation solution and a preparation method and application thereof. Background technique [0002] Adipose-derived stem cells (ADSCs) are a kind of stem cells with multi-directional differentiation potential isolated from adipose tissue in recent years. They have the characteristics of general stem cells such as rapid expansion and not easy to age. Human adipose stem cells, namely human adipose-derived mesenchymal stem cells, are a kind of adult stem cells widely used in the field of tissue engineering and regenerative medicine, which have the same multi-directional differentiation potential as bone marrow mesenchymal stem cells. ADSCs showed fibroblast-like growth, abundant cytoplasm and nucleoli, arranged in parallel or swirl-like arrangement. Cell cycle analysis showed that 69% of cells were in G0 / G1 phase, 24% in S phase, and 8% i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02C07K7/08
CPCA01N1/0221C07K7/08
Inventor 李倩
Owner 北京九思九如健康科技有限公司
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