Basic & Clinical Medicine ›› 2013, Vol. 33 ›› Issue (11): 1410-1417.

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Store bank building methods of human amniotic mesenchymal stem cells

  

  • Received:2013-08-26 Revised:2013-09-17 Online:2013-11-05 Published:2013-10-28

Abstract: Objective To build human amniotic mesenchymal stem cell bank for the application of MSCs provide sufficient source of cells. methods amniotic membrane were isolated from clean placenta,the human amniotic mesenchymal stem cell underwent primary culture and subculture from amniotic membrane. Cell growth pattern was detected by MTT assay. Flow cytometry was applied to analyze cell phenotype and stem cell markers. The different differential medium was used to detect its multi-directional differential ability. Such as Cell cycle, used identification of their biological characteristics. ofTransplantation of mesenchymal stem cells of magnetic nanoparticles markers under human amniotic membrane in mice of skin wound model to evaluate wound repair. results analysis shows that these amniotic -derived cells are adherent cells with typical fibroblast morphology. Cells extend not found in the process of bacteria, fungi, mold, mycoplasma, endotoxin pollution, etc. Flow cytometric analysis reveals that human amniotic mesenchymal stem cells are consistently positive for MSCs labeled with CD90 and CD105, and negative for the hematopoietic markers CD45, CD14, CD34 and HLA-DR. The cells can differentiate into osteoblasts and chondrogenic and adipogenesis under different conditioned mediumcultured conditions. Cell cycle analysis showed that most cells in G1 phase, only a few cells in the proliferation period G2. In vitro experiments show that human amniotic membrane mesenchymal stem cells can promote wound healing in mice, compared with the control group a very significant difference(p<0.05)Conclusion These results proved that these cells are human amniotic -derived MSCs. amniotic mesenchymal stem cells compared with bone marrow mesenchymal stem cells have a similar biological characteristics. Preliminary caracterizeed inventory cells biological characteristics and the established corresponding testing standards of hAMSCs in the quality control system.

Key words: Key words: human amniotic mesenchymal stem cells, biological characteristics, cell differentiation, surface marker , cell bank