兔骨髓基质干细胞的体外培养及生物学活性研究  

Biological activity of the cultured rabbit bone marrow-derived mesenchymal stem cells

作　　者：张晨[1] 王坤正[1] 强辉[1] 时志斌[1] 党晓谦[1] 黄向辉[2] 

机构地区：[1]西安交通大学医学院第二附属医院骨科,陕西西安710004 [2]陕西省人民医院骨科,陕西西安710068

出　　处：《西安交通大学学报（医学版）》2010年第2期138-142,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.30600624)~~

摘　　要：目的体外分离培养、鉴定兔骨髓基质干细胞(BMSCs)并探讨其分化潜能。方法应用密度梯度离心联合贴壁筛选的方法,体外分离培养兔骨髓来源的骨髓基质干细胞,培养过程中倒置显微镜下观察其生物学特性,HE染色和CD34、CD44免疫组织化学染色行细胞学鉴定;应用体外成骨与成脂诱导分化检测其生物学活性。结果体外实验成功分离培养兔BMSCs,HE染色显示细胞均一性好;CD44强阳性表达,CD34阴性表达。诱导成骨结果显示BMSCs具有较强的成骨活性,ALP染色阳性;成脂诱导结果显示BMSCs具有较强的成脂活性,油红"O"脂肪染色阳性。结论BMSCs体外具有潜在的多向分化能力。Objective To observe the effect of the in vitro isolation,culture and identification of rabbit bone marrow-derived mesenchymal stem cells（BMSCs） and explore the differentiation potentials of BMSCs.Methods BMSCs were isolated and purified by density gradient centrifugation combined with attachment culture method.The biological characteristics of BMSCs were observed under an inverted microscope.The BMSCs were identified with HE staining and CD34/CD44 immunohistochemical staining.The biological activity of BMSCs was detected by osteogenic induction and steatogenic induction.Results BMSCs were isolated successfully in vitro and had good homogenicity.Immunohistochemical staining of CD34/CD44 showed strong positive expression of CD44 and negative expression of CD34.Fortis osteogenic ability and positive staining of ALP could be detected by osteogenic induction,and fortis steatogenic ability and positive staining of axungia could be detected by steatogenic induction.Conclusion BMSCs have the capability of multi-directional differentiation in vitro.

关 键 词：骨髓基质干细胞 细胞培养 多向分化 

分 类 号：R318[医药卫生—生物医学工程] R392.11[医药卫生—基础医学]