Culture and identification of the human bone marrow mesenchymal stem cells in vitro
- VernacularTitle:人骨髓间充质干细胞体外培养及鉴定
- Author:
Xue LIANG
- Publication Type:Journal Article
- Keywords:
Human bone marrow mesenchymal stem cells;
Hematopoietic inductive microenvironment;
Cell culture
- From:
Journal of Medical Postgraduates
2003;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To isolate,culture and identify the human bone marrow mesenchymal stem cells(hBMSC). Methods:Human bone-marrow mononuclear cells(MNC) were isolated by centrifugation over Percoll(1.073 kg/L)and cultured in a BMSC culture system in vitro using their adherent characteristics.Many kinds of cellular molecules were measured to identify the cultured cells.Results:The cells show fibroblast-like irregular appearance,many cytoplasm ecptoma,puff and large nucleus with obvious nucleoli.The flow cytometry shows that 2.09% of the cells are CD34 positive,94.46% of the CD34 negative cells are CD29 positive,and CD54 and CD105 are possibly positive.The cell cycle observation and transmission electron microscopy show that the cells are in the primitive status.The immunocytochemistry detection shows that CD14,CD45 and collagen protein II are negative;CD106,CD166,fibronectin,vimentin and collagen fibers acidic protein(GFAP) are positive;staining for glycogen is strongly positive;staining for alkaline phosphatase is negative.The cells can be induced into osteoblast in proper inductive and differentiating culture circumstances.All the above results indicate that the isolation and culture of hBMSC are successful.Although the hBMSC can be serially subcultivated,the cells appear to be aging and they can not be kept by frozen. Conclusion:The steady ex vivo expansion system in which hBMSC can be obtained has been established.
