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Method for improving osteogenic differentiation efficiency of human amniotic mesenchymal stem cells and application thereof

A technology for stem cell and osteogenic differentiation, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve the problem of undiscovered human-derived factor stem cells, etc., to improve the efficiency of osteogenic differentiation and promote expression. and activity, the effect of avoiding the difficulty of osteoblasts

Inactive Publication Date: 2017-03-15
AFFILIATED HOSPITAL OF ZUNYI MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, so far, no human-derived factors have been discovered to regulate the differentiation of stem cells.

Method used

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  • Method for improving osteogenic differentiation efficiency of human amniotic mesenchymal stem cells and application thereof
  • Method for improving osteogenic differentiation efficiency of human amniotic mesenchymal stem cells and application thereof
  • Method for improving osteogenic differentiation efficiency of human amniotic mesenchymal stem cells and application thereof

Examples

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Embodiment 1

[0022] Example 1: Method and Application of Improving Osteogenic Differentiation Efficiency of Human Amniotic Mesenchymal Stem Cells

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

[0024] The amniotic membrane was bluntly removed from the fresh placenta of healthy full-term cesarean section, and the amniotic membrane was washed with D-Hank's solution containing 1% double antibody (final concentration: penicillin: 100 IU / mL, streptomycin: 100 IU / mL, fresh before use). Preparation) repeatedly rinse the amniotic membrane to remove residual blood stains and surface mucus. Cut the amniotic membrane into 1-3 cm 2 Divide into 50 mL 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 ℃ at 150-185rpm for about 30 minutes , after filtering through a 300-mesh stainless steel filter, discard the digestion solution containing amniot...

Embodiment 2

[0031] Example 2. The use of human amniotic mesenchymal stem cells to promote osteogenesis in other aspects

[0032] Human amniotic mesenchymal stem cells are induced to differentiate into osteoblasts in vitro by the induction composition of the present invention, and can be used as seed cells for bone tissue engineering and cell transplantation. It has a certain preventive effect on fractures and bone metabolic diseases. Using the induction composition of the present invention to induce differentiation of human amniotic mesenchymal stem cells as seed cells to replace osteoblasts can avoid the problems of difficult acquisition of osteoblasts in vitro and high immunogenicity.

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Abstract

The invention relates to a method for improving osteogenic differentiation efficiency of human amniotic mesenchymal stem cells. The method adds an induction composition in the human amniotic mesenchymal stem cells, wherein the composition comprises 0.1-2g / L of hyaluronic acid, 0-200mg / L of prostaglandin receptor-2 selective agonist, 1-100nmol / L of dexamethasone, 0-10nmol / L of beta-glycerophosphate and 1-100mg / L of ascorbic acid. Compared with a conventional inducing method, the osteogenic differentiation efficiency of the human amniotic mesenchymal stem cells can be significantly improved, expression and activity of alkaline phosphatase are promoted, and expression of osteogenesis-related genes of RunX2, Osx, BSP, Ocn, ALP, Col1a1 and the like and formation of mineralized calcium nodules are enhanced. The method for improving the osteogenic differentiation efficiency of the human amniotic mesenchymal stem cells can be applied in osteogenic differentiation of the human mesenchymal stem cells, and applied in bone diseases of bone defects, bone trauma and the like.

Description

technical field [0001] The invention belongs to the technical field of stem cell biotechnology and bone tissue engineering, and relates to a method and application for improving the osteogenic differentiation efficiency of human amniotic mesenchymal stem cells. Background technique [0002] Bone damage or defect due to trauma, infection, degenerative disease, tumor resection and other reasons is a common and frequently-occurring disease in orthopedic diseases. Taking osteoarthritis as an example, there are currently about 150 million patients in my country. WHO estimates that by 2025, there will be more than 800 million patients worldwide. The number is second only to cardiovascular-related diseases, and it has become an important disease that seriously affects human health. At present, the repair and functional reconstruction of bone defects has become a major problem in the medical field because traditional treatment methods are difficult to achieve ideal results. In rece...

Claims

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Application Information

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IPC IPC(8): C12N5/077C12N5/0775A61L27/38
CPCA61L27/3834A61L27/3847A61L2430/02C12N5/0654C12N2500/34C12N2500/38C12N2500/42C12N2501/30C12N2501/905C12N2506/1392
Inventor 肖建辉张令涛刘如明
Owner AFFILIATED HOSPITAL OF ZUNYI MEDICAL COLLEGE
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