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Mesenchymal stem cell in-vitro culture kit and application thereof

A stem cell and in vitro culture technology, applied in the field of cell biology, can solve the problems of difficult quality control in the preparation process, decline in proliferation and differentiation ability, and reduction in the number of stem cells, and achieve high success rate, fast cell crawling, and lower production costs. Effect

Pending Publication Date: 2022-03-11
深圳市申友健康管理有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, bone marrow-derived mesenchymal stem cells have the following problems: with aging, the number of stem cells decreases significantly, and the ability to proliferate and differentiate significantly declines; the preparation process is not easy to quality control; transplantation to allogeneic cells may cause immune reactions; There are injuries, etc., which limit its application in clinical treatment

Method used

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  • Mesenchymal stem cell in-vitro culture kit and application thereof
  • Mesenchymal stem cell in-vitro culture kit and application thereof
  • Mesenchymal stem cell in-vitro culture kit and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0065] This embodiment provides a kit for culturing mesenchymal stem cells in vitro, the kit for culturing mesenchymal stem cells in vitro includes an attachment-promoting medium, a regulating medium and a proliferation-promoting medium;

[0066] The medium for promoting adhesion includes cell culture supplement 1% (v / v), fetal bovine serum 1% (v / v), rhGM-CSF800IU / mL, bFGF 80ng / mL, SCF 80ng / mL and IL-3 80ng / mL, the balance is stem cell culture medium;

[0067] The conditioned medium includes cell culture supplement 1% (v / v), TPO 60IU / mL, SCF 60ng / mL and IL-3 60ng / mL, and the balance is stem cell culture medium;

[0068] The proliferation-promoting medium includes cell culture supplement 1% (v / v), TPO 50IU / mL, SCF 50ng / mL and IL-3 50ng / mL, and the balance is stem cell medium.

Embodiment 2

[0070] This embodiment provides a kit for culturing mesenchymal stem cells in vitro, the kit for culturing mesenchymal stem cells in vitro includes an attachment-promoting medium, a regulating medium and a proliferation-promoting medium;

[0071] The adhesion-promoting medium includes cell culture supplement 6% (v / v), fetal calf serum 6% (v / v), rhGM-CSF1000IU / mL, bFGF 100ng / mL, SCF 100ng / mL and IL-3 100ng / mL, the balance is stem cell culture medium;

[0072] The conditioned medium includes cell culture supplement 6% (v / v), TPO 90IU / mL, SCF 90ng / mL and IL-3 90ng / mL, and the balance is stem cell culture medium;

[0073] The proliferation-promoting medium includes cell culture supplement 6% (v / v), TPO 70IU / mL, SCF 70ng / mL and IL-3 70ng / mL, and the balance is stem cell medium.

Embodiment 3

[0075] This embodiment provides a kit for culturing mesenchymal stem cells in vitro, the kit for culturing mesenchymal stem cells in vitro includes an attachment-promoting medium, a regulating medium and a proliferation-promoting medium;

[0076] The medium for promoting adhesion includes cell culture supplement 10% (v / v), fetal calf serum 10% (v / v), rhGM-CSF 1200IU / mL, bFGF 120ng / mL, SCF 120ng / mL and IL- 3 120ng / mL, the balance is stem cell culture medium;

[0077] The conditioning medium includes cell culture supplement 10% (v / v), TPO 100IU / mL, SCF 100ng / mL and IL-3 100ng / mL, and the balance is stem cell culture medium;

[0078] The proliferation-promoting medium includes cell culture supplement 10% (v / v), TPO 90IU / mL, SCF 90ng / mL and IL-3 90ng / mL, and the balance is stem cell medium.

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Abstract

The invention provides a mesenchymal stem cell in-vitro culture kit and application thereof. The mesenchymal stem cell in-vitro culture kit comprises an adherence-promoting culture medium, a regulating culture medium and a proliferation-promoting culture medium, the adherence promoting culture medium comprises a stem cell culture medium, a cell culture supplement, fetal calf serum, adherence promoting cell factors and IL-3; the regulating culture medium comprises a stem cell culture medium, a cell culture supplement, a regulating cell factor and IL-3; the proliferation-promoting culture medium comprises a stem cell culture medium, a cell culture supplement, proliferation-promoting cell factors and IL-3. The invention further provides an in-vitro culture method of the mesenchymal stem cells and the mesenchymal stem cells cultured by the in-vitro culture method. According to the invention, the optimized culture medium is matched with the culture method, so that the mesenchymal stem cells with strong division ability and high purity can be rapidly obtained, and the application prospect is wide.

Description

technical field [0001] The invention belongs to the technical field of cell biology, and in particular relates to a mesenchymal stem cell in vitro culture kit and application thereof. Background technique [0002] Mesenchymal stem cells are a kind of pluripotent stem cells with the ability of self-renewal and multilineage differentiation. Mesenchymal stem cells were first discovered in the bone marrow, and later found that they also exist in various tissues during human development and development, including bone marrow, fat, synovium, bone, muscle, lung, liver, pancreas, amniotic fluid and umbilical cord blood Wait. [0003] Mesenchymal stem cells have been widely used in clinical research. In addition to promoting recovery of hematopoiesis and co-transplantation with hematopoietic stem cells to treat leukemia and refractory anemia, etc., they are also used for cardiovascular and cerebrovascular diseases, liver cirrhosis, bone and muscle degeneration In the research on th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0668C12N2501/20C12N2501/2303C12N2501/22C12N2501/115C12N2501/125
Inventor 易亮梁秋彬罗国辉钱春明颜春江汪金球黄景云邵小兵褚志伟吴佑星
Owner 深圳市申友健康管理有限公司
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