脐带间充质干细胞的体外分离及生物学特性观察(英文)  

Isolation and biological characteristics of umbilical cord mesenchymal stem cells

作　　者：田新[1] 韦红艳[1] 符仁义[1] 

机构地区：[1]广西医科大学第四附属医院儿科,广西壮族自治区柳州市545005

出　　处：《中国组织工程研究与临床康复》2009年第32期6340-6344,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:脐带组织依赖于母体免疫系统的保护,且胚胎自身免疫系统相对不发育、MHC表达低下,故来源于脐带的间充质干细胞的生物学特性已成为目前研究热点。目的:拟在体外分离培养人脐带组织间充质干细胞,并观察其多向分化能力。设计、时间及地点:细胞学体外观察,于2006-10/2007-05在四川大学组织工程重建实验室完成。材料:脐带来源于胎龄37～40周的健康新生儿,由四川大学华西第二医院产房提供。方法:取脐带沿血管长轴切开,去掉血管,再将脐带重新缝合形成环状,灌入胶原酶悬液,6～8h后灌洗离心,获取细胞后贴壁法分离培养、扩增,细胞呈集落生长后传代。取传至第5代细胞,分别加入成骨、成脂诱导分化培养基。主要观察指标:脐带间充质干细胞的形态、生长状况、表面抗原分子的表达、多向分化潜能。结果:去除血管后获取脐带组织细胞的方法可获得贴壁生长的细胞,呈短棒状或梭形样细胞,易扩增和形成集落;高表达基质细胞抗原CD29,CD51,CD71,而不表达CD34,CD45及HLA-DR等造血干细胞抗原分子;成骨诱导后茜素红染色胞浆中有大量的钙沉积,碱性磷酸酶钙钴法染色胞质呈灰黑色,阳性细胞率>85%;成脂诱导后油红O染色示胞浆充满油滴空泡。结论:脐带组织存在具有分化能力的间充质干细胞,并可在体外进行培养扩增形成集落细胞传代,传代细胞表达基质细胞表面抗原,能够向成骨细胞、成脂肪细胞方向分化。BACKGROUND： Cord tissues, which immune system is relatively non-development with low expression of MHC, rely on the protection of the mother＇s immune system. Accordingly, studies underlying biological characteristics of umbilical cord mesenchymal stem cells （MSCs） arouse more and more attention. OBJECTIVE： To isolate, separate, and culture MSCs from the tissues of umbilical cord, and to explore the ability of multi-directional differentiation. DESIGN, TIME AND SETTING： The in vitro cytology experiment was performed at the Laboratory of Tissue Engineering of Sichuan University from October 2006 to May 2007. MATERIALS： The umbilical cord of healthy neonate with 37-40 weeks gestational age was obtained from delivery room of the Second West China Hospital, Sichuan University. METHODS： Pieces of umbilical cord were dissected along its long axis, vessel was pulled away, then cord was tied together with a suture creating ＂loops＂, then it was placed into collagenase solution, after 6-8 hours, suspended cells obtained from the loops suspension were centrifuged and cultured with adherent method, passaged when single call-derived colonies were appeared. Finally, the 5^th generation of cells was induced differentiation towards osteoblasts and lipoblasts. MAIN OUTCOME MEASURES： Morphology, growth, surface antigen expression, as well as the multiple differentiation potential of umbilical cord MSCs. RESULTS： The adherent cells obtained from umbilical cord after removal vessels showed short rod-like or fusiform shape, which can be proliferated and form single cell-derived colonies. Cells isolated from the umbilical cord high expressed integrin markers （CD29, CD51, CD71） but not hematopoietic lineage markers （CD34, CD45 and HLA-DR）. These colonies can differentiate into osteoblasts that produce mineralized matrices, stained by alizarin red and alkaline phosphatase, and differentiate adipocytes that accumulate lipid vacuoles and are demonstrated by morphology, stained by oil red. CONCLUSION： Umbil

关 键 词：脐带 间充质干细胞 分化 

分 类 号：R394.2[医药卫生—医学遗传学]