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Recombinant mesenchymal stem cells and preparing method thereof

A mesenchymal stem cell and cell technology, applied in the field of recombinant mesenchymal stem cells and their preparation, can solve problems affecting the effect of stem cells in repairing local damaged tissues, and achieve the effects of increasing migration speed and distance, enhancing differentiation potential, and increasing the number

Active Publication Date: 2017-11-10
SUZHOU UNIV
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  • Description
  • Claims
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Problems solved by technology

For example, it has been reported in the literature that after 26 days of spinal cord injury surgery, less than 1% of the transplanted cells can be detected in the damaged area, which seriously affects the effect of stem cells in repairing the local damaged tissue.

Method used

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  • Recombinant mesenchymal stem cells and preparing method thereof
  • Recombinant mesenchymal stem cells and preparing method thereof
  • Recombinant mesenchymal stem cells and preparing method thereof

Examples

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Embodiment 1

[0030] Example 1 Preparation and Verification of Recombinant Mesenchymal Stem Cells

[0031] (1) Isolation, primary and subculture of mesenchymal stem cells, the specific methods are as follows:

[0032] Primary cells were extracted from rat mesenchymal stem cells by the whole bone marrow adherent culture method, using low-sugar (L-DMEM) medium containing 10% serum, and placed in a 37°C incubator containing 5% carbon dioxide.

[0033] (2) Stem cell construction:

[0034] Transfect miR-124 or miR-21 into mesenchymal stem cells by liposome transfection to construct recombinant mesenchymal stem cells that highly express miR-124 or miR-21. The specific operations are as follows:

[0035] 1. Divide MSCs in 5×10 4 / ml density in 35mm cell culture dish. Transfection was started after the cells grew to 80% confluence. Ensure that the final concentration of transfection is 50nM miRNA promoter.

[0036] 2. Add 5ul pre-configured mimics solution to 245μl L-DMEM, mix gently, and let ...

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Abstract

The invention relates to recombinant mesenchymal stem cells in which miR-124 and miR-21 are transfected, and further provides a preparing method of the recombinant mesenchymal stem cells. The method includes the steps of transfecting miR-124 and miR-21 in mesenchymal stem cells, and screening out the cells in ascended expression level to obtain the recombinant mesenchymal stem cells. The transfer ability and proliferation ability of the recombinant mesenchymal stem cells are both improved, and therefore the treatment effect for treating diseases of a central nervous system and the like through MSCs cell transplantation is improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a recombinant mesenchymal stem cell and a preparation method thereof. Background technique [0002] Mesenchymal stem cells (MSCs) are mesoderm-derived adult stem cells with multi-differentiation potential, which exist in connective tissue and organ interstitium. MSCs come from a wide range of sources, including bone marrow, fat, umbilical cord (blood) , gums, skeletal muscle and other tissues, among which the bone marrow tissue is the most abundant. MSCs have the unique unlimited proliferation and self-renewal ability of stem cells, and can still maintain their "stemness" after in vitro subculture and cryopreservation recovery; they have multi-directional differentiation potential, and can differentiate into bone cells under suitable in vivo and in vitro environments , chondrocytes, adipocytes, nerve cells and other cell types; with low immunogenicity, it inhibits the proliferation ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12N15/88
CPCC12N5/0663C12N15/88C12N2510/00
Inventor 曲静刘衍
Owner SUZHOU UNIV
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