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Method for promoting differentiation of human amniotic mesenchymal stem cells into chondrocytes and application thereof

A technology of mesenchymal stem cells and chondrocytes, which is applied in the field of promoting the differentiation of human amniotic mesenchymal stem cells into chondrocytes, can solve the problems of no obvious change, dependence on promotion of proliferation, etc., and achieve the effect of avoiding the problem of high immunogenicity

Inactive Publication Date: 2017-01-04
AFFILIATED HOSPITAL OF ZUNYI MEDICAL COLLEGE
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Problems solved by technology

The results suggest that low molecular weight HA can promote cell proliferation, and dose-dependently enhance the expression of osteocalcin gene mRNA, a specific marker of osteoblast differentiation, but have no significant changes in alkaline phosphatase, bone formation and other indicators, while the other two The molecular weight of HA can significantly promote the proliferation of osteoprogenitor cells and induce differentiation into osteocytes. Kimata et al. observed that HA regulates the proliferation of mouse epidermal fibroblasts in the logarithmic growth phase, and found that there are positive and negative effect regulation phenomena, which depend entirely on the addition of The cell density in a certain dose range of HA, when cultured at a relatively high density, showed a dose-dependent tendency to promote proliferation, and vice versa, inhibited proliferation

Method used

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  • Method for promoting differentiation of human amniotic mesenchymal stem cells into chondrocytes and application thereof
  • Method for promoting differentiation of human amniotic mesenchymal stem cells into chondrocytes and application thereof
  • Method for promoting differentiation of human amniotic mesenchymal stem cells into chondrocytes and application thereof

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Embodiment 1

[0022] Example 1: The method and application of promoting the differentiation efficiency of human amniotic mesenchymal stem cells into chondrocytes

[0023] 1. Isolation and culture of human amniotic mesenchymal stem cells

[0024] The fresh placenta of healthy full-term cesarean section was bluntly mechanically stripped the amniotic membrane, and the D-Hank's solution containing 1% double antibody (final concentration of penicillin: 100 IU / mL, streptomycin: 100 IU / mL, freshly prepared before use) ) Wash the amniotic membrane repeatedly to remove residual blood stains and surface mucus. Cut the amniotic membrane into 1-3 cm 2Divide into 50mL centrifuge tubes, add 0.05% trypsin digestion solution containing 0.02% EDTA-2Na about 2 times the volume of amnion tissue, and digest in a constant temperature water bath at 35-37°C at 150-185rpm for about 30 minutes. After filtering through a 300-mesh stainless steel filter, discard the digestion solution containing amniotic epithelial...

Embodiment 2

[0033] Example 2. Promoting the use of human amniotic mesenchymal stem cell chondrocytes in other aspects

[0034] The human amniotic mesenchymal stem cells are induced to differentiate into chondrocytes by using the inducing composition of the present invention in vitro, and can be used as seed cells for cartilage tissue engineering and cell transplantation. It has a certain preventive effect on cartilage damage diseases. Using the induction composition of the present invention to induce differentiation of human amniotic mesenchymal stem cells as seed cells to replace chondrocytes can avoid the problems of difficult acquisition and high immunogenicity of chondrocytes in vitro.

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Abstract

The invention relates to a method for promoting differentiation of human amniotic mesenchymal stem cells into chondrocytes. The method comprises the step of adding an induction composition into the human amniotic mesenchymal stem cells, wherein the induction composition comprises 0.01-2g / L of hyaluronic acid and 0-100mg / L of ascorbic acid. Compared with the conventional induction method, the method disclosed by the invention has the advantages that the effect of promoting differentiation of human amniotic mesenchymal stem cells into chondrocytes can be obviously improved, glycosaminoglycan and type II collagen expression is strongly positive, expression of chondrogenesis related genes and proteins such as Col2alpha1, ACAN, Sox9 and the like is obviously improved, and the efficiency of differentiating the human amniotic mesenchymal stem cells into chondrocytes can be obviously improved. The induction composition disclosed by the invention can be used for differentiation of human amniotic mesenchymal stem cells into chondrocytes and cartilage injury diseases.

Description

technical field [0001] The invention belongs to the technical field of stem cell biotechnology and bone tissue engineering, and particularly relates to a method and application for promoting the differentiation of human amniotic mesenchymal stem cells into chondrocytes. Background technique [0002] Articular cartilage injury caused by sports trauma, degenerative disease, infection, tumor resection, etc. is a common and frequently-occurring disease in orthopedic diseases, and has become an important disease affecting human health. Since articular cartilage has no blood supply, and chondrocytes are embedded in a thick extracellular matrix, it is difficult to migrate to the damaged site to participate in repair. However, traditional treatment methods have extremely limited efficacy. Articular cartilage defect has always been a difficult point in the treatment of orthopedic clinical diseases and an important topic of research. At present, the methods for treating cartilage in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077C12N5/0775A61L27/38
CPCC12N5/0655A61L27/3817A61L2430/06C12N2500/38C12N2501/905C12N2506/1392C12N2509/00
Inventor 肖建辉张清芳刘如明
Owner AFFILIATED HOSPITAL OF ZUNYI MEDICAL COLLEGE
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