全骨髓贴壁法分离培养人骨髓间充质干细胞及成骨诱导分化  

Isolation,culture and osteogenic differentiation of human bone marrow stem cells by using whole bone marrow adherence method

作　　者：张利铭[1,2] 陈智超[1] 邹萍[1] 李秋柏[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院血液科,武汉430022 [2]华中科技大学同济医学院附属荆州医院血液科

出　　处：《临床血液学杂志》2012年第3期317-319,共3页Journal of Clinical Hematology

基　　金：国家自然科学基金资助项目(No:81071943)

摘　　要：目的:探讨体外分离骨髓间充质干细胞的方法及其成骨潜能。方法:应用全骨髓贴壁法分离培养人骨髓间充质干细胞,倒置显微镜下观察细胞形态,流式细胞仪检测细胞周期及免疫表型,碱性磷酸酶及茜素红染色检测成骨能力。结果:传代后的人骨髓间充质干细胞形态均一呈成纤维细胞样。第三代骨髓间充质干细胞免疫表型CD90、CD13、CD105、CD44阳性,CD34、CD45、CD14阴性。96.47%细胞处于G0/G1期,14d、21d成骨诱导培养后碱性磷酸酶及茜素红染色阳性。结论:应用全骨髓贴壁法体外分离培养人骨髓间充质干细胞简便、经济、高效。Objective：To explore a better method of isolation human bone marrow stem cells and study its potentiality of osteogenic differentiation in vitro. Method：Human bone marrow stem cells were isolated and cultured by using whole bone marrow adherence method.The cell morphology was observed under the inverted phase contrast microscope.The cell cycle and immunological phenotype were examined by flow cytometry.Osteogenic ability was examined by alkaline phosphatase（ALP） and alizarin red stain. Result：The passage cells were homogeneous in shape,similar to fibroblasts.The passage 3 cells were postive for CD90,CD13,CD105,CD44,but negative for CD34,CD45,CD14,and the overwhelming majority of cells were in G0/G1 phase.Following by 14,21 days of osteogenic differentiation,the ALP and alizarin red stains were postive in osteoblasts. Conclusion：Whole bone marrow adherence method as a simple,cheap and high efficiency performance is a good culture method for human bone marrow stem cells.

关 键 词：间充质干细胞 全骨髓贴壁法 细胞培养 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]