中国组织工程研究2012，Vol.16Issue(14)：2515-2519,5.DOI:10.3969/j.issn.1673-8225.2012.14.009
人骨髓间充质干细胞的体外培养、鉴定及成骨分化
In vitro culture, identification and osteogenic differentiation of human bone marrow mesenchymal stem cells
刘伟 1刘萌 2祝劲松 1郝红伟 1董宁征 2周海斌1
作者信息
- 1. 苏州大学附属第二医院骨科,江苏省苏州市215004
- 2. 苏州大学唐仲英血液学研究中心,江苏省苏州市,215004
- 折叠
摘要
Abstract
BACKGROUND: Research approaches and measurement index regarding osteogenic differentiation of bone marrow mesenchymal stem cells in vitro are not fully considered at home and abroadOBJECTIVE: To establish and consummate a set of methods of isolating, culturing and identifying human bone marrow-derived mesenchymal stem cells (hBMSCs) and investigate their osteogenic differentiation ability in vitroMETHODS: hBMSCs were isolated using the method of density gradient centrifugation The cell surface phenotypes of BMSCs were identified by flow cytometry Passage 3 hBMSCs were induced by osteogemc culture medium for 2-3 weeks RESULTS AND CONCLUSION: The primary cultured hBMSCs grew and proliferated vigorously after subculture Passage 3 hBMSCs were positive for CD44, CD73, CD90 and were negative for CD34 After osteogemc induction, alkaline phosphatase activity was increased and cells were stained positive by Gomori, Von kossa and alizarin red RT-PCR results showed the gene expression of collagen I, alkaline phosphatase, osteocalcin, bone sialoprotein, osteopotin and osteonectin This confirms that the primary hBMSCs can differentiate into osteoblasts in vitro successfully These findings suggest that a stable, mature method of isolating, cultunng and amplifying hBMSCs has been established.关键词
骨髓间充质干细胞/骨髓/分离培养/成骨细胞/成骨分化分类
医药卫生引用本文复制引用
刘伟,刘萌,祝劲松,郝红伟,董宁征,周海斌..人骨髓间充质干细胞的体外培养、鉴定及成骨分化[J].中国组织工程研究,2012,16(14):2515-2519,5.
