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Method for stimulating cartilage tissue differentiation of mesenchymal stem cells

A technology of mesenchymal stem cells and cartilage tissue, applied in the field of stimulating cartilage tissue differentiation of mesenchymal stem cells, can solve the problems of long differentiation cycle, low expression efficiency, and poor effect, so as to promote cartilage repair ability, improve expression efficiency, and reduce cycle effect

Pending Publication Date: 2021-11-09
THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In order to solve the above-mentioned problems, the object of the present invention is to provide a method for stimulating cartilage tissue differentiation of mesenchymal stem cells, which solves the long cycle of cartilage tissue differentiation of mesenchymal stem cells in the prior art, low expression efficiency and the lack of efficacy in the treatment of arthritis. and ineffectiveness in promoting cartilage repair capacity

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  • Method for stimulating cartilage tissue differentiation of mesenchymal stem cells
  • Method for stimulating cartilage tissue differentiation of mesenchymal stem cells
  • Method for stimulating cartilage tissue differentiation of mesenchymal stem cells

Examples

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Effect test

Embodiment 1

[0033] A method for stimulating cartilage tissue differentiation of mesenchymal stem cells provided in Example 1 of the present invention is specifically (such as figure 1 shown):

[0034] S1. Adding dendritic cells isolated and cultured in vitro to TGF-β3 at a concentration of 10 ng / mL and IFN1&2 at a concentration of 20 ng / mL to obtain plasmacytoid dendritic cells; wherein, the dendritic cells in vitro The isolation and culture method is as follows: the hematopoietic progenitor cells are placed in DMEM solution including 10% fetal bovine serum, 1% penicillin / streptomycin, 20ng / mL GM-CSF and 20ng / mLIL-4, cultured for 5 days, collected and cultured resulting dendritic cells;

[0035] S2. When the number of plasmacytoid dendritic cells reaches 1.0×10 5 At one time, the expression of tolerance in plasmacytoid dendritic cells was induced by indoleamine-2,3-dioxygenase-1 at a concentration of 10 ng / mL, and the induction time was 3 days to obtain tolerance plasma Cell-like dendr...

Embodiment 2

[0039] A method for stimulating cartilage tissue differentiation of mesenchymal stem cells provided in Example 2 of the present invention is specifically:

[0040]S1. Adding dendritic cells isolated and cultured in vitro to TGF-β3 at a concentration of 8 ng / mL and IFN1&2 at a concentration of 18 ng / mL to obtain plasmacytoid dendritic cells; wherein, the dendritic cells in vitro The isolation and culture method is as follows: the hematopoietic progenitor cells are placed in DMEM solution including 8% fetal bovine serum, 0.8% penicillin / streptomycin, 18ng / mL GM-CSF and 18ng / mLIL-4, cultured for 4 days, collected and cultured resulting dendritic cells;

[0041] S2. When the number of plasmacytoid dendritic cells reaches 1.0×10 5 At one time, the expression of tolerance in plasmacytoid dendritic cells was induced by indoleamine-2,3-dioxygenase-1 at a concentration of 8 ng / mL, and the induction time was 3 days to obtain tolerance plasma Cell-like dendritic cells;

[0042] S3, pu...

Embodiment 3

[0044] A method for stimulating cartilage tissue differentiation of mesenchymal stem cells provided in Example 3 of the present invention is specifically:

[0045] S1. Adding dendritic cells isolated and cultured in vitro to TGF-β3 at a concentration of 12 ng / mL and IFN1&2 at a concentration of 22 ng / mL to obtain plasmacytoid dendritic cells; wherein, the dendritic cells in vitro The isolation and culture method is as follows: the hematopoietic progenitor cells are placed in DMEM solution including 12% fetal bovine serum, 1.2% penicillin / streptomycin, 22ng / mL GM-CSF and 22ng / mL IL-4, cultured for 6 days, collected cultured dendritic cells;

[0046] S2. When the number of plasmacytoid dendritic cells reaches 1.0×10 5 At one time, the expression of tolerance in plasmacytoid dendritic cells was induced by indoleamine-2,3-dioxygenase-1 at a concentration of 12 ng / mL, and the induction time was 3 days to obtain tolerance plasma Cell-like dendritic cells;

[0047] S3, put 1x10 6...

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Abstract

The invention discloses a method for stimulating cartilage tissue differentiation of mesenchymal stem cells, which comprises the following steps: 1) adding dendritic cells separated and cultured in vitro into TGF-beta3 and IFN1 and 2 to obtain plasma cell-like dendritic cells; (2) when the number of the plasma cell-like dendritic cells reaches a certain number, inducing tolerant expression of the plasma cell-like dendritic cells by adopting indoleamine-2, 3-dioxygenase-1, so as to obtain tolerant plasma cell-like dendritic cells; and (3) co-culturing the tolerant plasma cell-like dendritic cells and the mesenchymal stem cells so as to achieve cartilage differentiation of the mesenchymal stem cells. According to the method, the tolerant plasma cell-like dendritic cells are adopted to stimulate the cartilage tissue differentiation of the mesenchymal stem cells, so that the cartilage tissue differentiation period of the mesenchymal stem cells is effectively shortened, the expression efficiency of proteins is also improved, and meanwhile, a solid foundation is laid for treating arthritis and promoting cartilage repair capability.

Description

technical field [0001] The invention belongs to the technical field of cartilage tissue differentiation, and in particular relates to a method for stimulating cartilage tissue differentiation of mesenchymal stem cells. Background technique [0002] Since chondrocytes are terminally differentiated cells with limited self-repair ability, the treatment of cartilage defects caused by trauma or osteoarthritis has always been a difficult problem in orthopedics. In recent years, with the development of cell transplantation or cartilage tissue engineering technology, the treatment of cartilage defects has made some progress. Autologous chondrocyte transplantation is currently recognized as a more effective method for the treatment of articular cartilage defects. However, autologous chondrocyte transplantation also has many disadvantages: ①The source of autologous chondrocytes is difficult. Since autologous chondrocyte transplantation needs to take a certain size of cartilage block...

Claims

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Application Information

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IPC IPC(8): C12N5/077C12N5/0775C12N5/0784
Inventor 阿哈得李文翠张辉王大平黄榕祥邓志钦
Owner THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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