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A kind of preparation method of human mesenchymal stem cell freeze-dried powder

A technology of stem cells and freeze-dried powder, applied in the field of stem cells, can solve the problems of allergy and rejection, the active components of stem cells cannot be kept intact, cytotoxic side effects, etc., to prevent cell damage and death, and the system components are simple and clear. good repeatability

Active Publication Date: 2020-03-24
北京殷氏细胞生物科技集团有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The present invention proposes a preparation method of human mesenchymal stem cell freeze-dried powder, which solves the cytotoxic side effects existing in the cell freezing technology in the prior art, the allergy and rejection that may be caused by heterologous serum, the nonviable stem cells and stem cells after freezing The problem that the active ingredients in the

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  • A kind of preparation method of human mesenchymal stem cell freeze-dried powder
  • A kind of preparation method of human mesenchymal stem cell freeze-dried powder
  • A kind of preparation method of human mesenchymal stem cell freeze-dried powder

Examples

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Embodiment 1

[0042] A preparation method of human mesenchymal stem cell freeze-dried powder, comprising the following steps:

[0043] 1) Collection: collect high-quality human mesenchymal stem cells with a passage number between 2 and 8 generations;

[0044] 2) Cultivation of mesenchymal stem cells: using a serum-free medium to amplify and passage human mesenchymal stem cells; the serum-free medium uses the medium with the application number CN2017100038074;

[0045] 3) Cryopreservation of mesenchymal stem cells: using a cell cryopreservation solution to freeze passaged human mesenchymal stem cells to -80 degrees; the cell cryopreservation solution contains active ingredients and their contents are as follows:

[0046] serial number active ingredient content(%) 1 Trehalose 0.1g / 100ml 2 Dextran 0.1g / 100ml 3 polyarginine 0.1mg / 100ml 4 Mannitol 0.1g / 100ml 5 Hydroxyethyl starch 0.1g / 100ml 6 Poloxamer (P188) 0.1g / 100ml 7 Glycer...

Embodiment 2

[0053] A preparation method of human mesenchymal stem cell freeze-dried powder, comprising the following steps:

[0054] 1) Collection: collect high-quality human mesenchymal stem cells with a passage number between 2 and 8 generations;

[0055] 2) Cultivation of mesenchymal stem cells: using a serum-free medium to amplify and passage human mesenchymal stem cells; the serum-free medium uses the medium with the application number CN2017100038074;

[0056] 3) Cryopreservation of mesenchymal stem cells: using a cell cryopreservation solution to freeze passaged human mesenchymal stem cells to -80 degrees; the cell cryopreservation solution contains active ingredients and their contents are as follows:

[0057] serial number active ingredient content(%) 1 Trehalose 9g / 100ml 2 Dextran 15g / 100ml 3 polyarginine 5mg / 100ml 4 Mannitol 5g / 100ml 5 Hydroxyethyl starch 5g / 100ml 6 Poloxamer (P188) 3g / 100ml 7 Glycerol 4g / 100...

Embodiment 3

[0064] A preparation method of human mesenchymal stem cell freeze-dried powder, comprising the following steps:

[0065] 1) Collection: collect high-quality human mesenchymal stem cells with a passage number between 2 and 8 generations;

[0066] 2) Cultivation of mesenchymal stem cells: using a serum-free medium to amplify and passage human mesenchymal stem cells; the serum-free medium uses the medium with the application number CN2017100038074;

[0067] 3) Cryopreservation of mesenchymal stem cells: using a cell cryopreservation solution to freeze passaged human mesenchymal stem cells to -80 degrees; the cell cryopreservation solution contains active ingredients and their contents are as follows:

[0068]

[0069]

[0070] The rest are water for injection and serum-free basal medium, and the basal medium is formed by mixing DMEM and F12 uniformly in a solution volume ratio of 1:1;

[0071] 4) Preparation of freeze-dried powder: prepare the human mesenchymal stem cells ...

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Abstract

The invention provides a method for preparing freeze-dried powder of human mesenchymal stem cells. The method includes the steps of 1) collecting, 2) mesenchymal stem cell culture, 3) mesenchymal stem cell cryopreservation, 4) freeze-dried powder preparation and 5) reactivation liquid addition. The freeze-dried powder prepared according to the method can be quickly dissolved and reactivate more than 60% of the stem cells after the reactivation liquid is added, and can retain cell factors, stem cell membranes and other special components with biological activity in the human mesenchymal stem cells effectively, so that a bottleneck that storage life of the active components of the stem cells is solved effectively. The method for preparing the freeze-dried powder of the human mesenchymal stem cells solves the problems that cell toxic and side effects exist, heterogenous sera possibly lead to allergies and rejection, no viable stem cells exist after freezing and the active components in the dry cells cannot be completely retained in a cell freezing technology in the prior art.

Description

technical field [0001] The invention relates to the technical field of stem cells, in particular to a preparation method of human mesenchymal stem cell freeze-dried powder. Background technique [0002] So far, the stem cells used in the treatment of human diseases and cosmetics in the world are mainly hematopoietic stem cells and mesenchymal stem cells, the latter mainly derived from placenta / umbilical cord, bone marrow and adipose tissue. Mesenchymal stem cells were originally discovered in the bone marrow. With their high self-renewal and multi-directional differentiation potential, mesenchymal stem cells are easy to isolate, culture and expand, have no tumorigenicity and ethical issues, and are easy to transfect and express foreign genes. , easy to obtain, transplant and avoid rejection and many other advantages, it has gradually become the focus of stem cell research, and has important research and application prospects, especially in cell therapy and gene therapy. It h...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775A61K8/98A61K8/81A61K8/73A61K8/67A61K35/28A01N1/02A61Q19/08
CPCA01N1/0226A61K8/676A61K8/735A61K8/8176A61K8/981A61K35/28A61Q19/08C12N5/0668C12N2500/34C12N2500/38C12N2511/00
Inventor 黄兵殷勤伟
Owner 北京殷氏细胞生物科技集团有限公司
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