贴壁法体外培养大鼠骨髓间质干细胞的生物学特点  

Biological characteristics of mesenchymal stem cells of rats cultured in vitro with adherent method

作　　者：冯智英[1] 张毅[1] 秦洁行[1] 苗玲[1] 

机构地区：[1]上海第二医科大学附属仁济医院神经内科,上海市200001

出　　处：《中国临床康复》2006年第9期37-39,i0002,共4页Chinese Journal of Clinical Rehabilitation

摘　　要：目的:观察大鼠骨髓间质干细胞在体外的提取、分离、培养和传代的特征,为进一步的研究工作提供实验基础。方法:实验于2004-06/12在上海第二医科大学附属仁济医院神经病学实验室完成。选取幼年雄性SD大鼠,处死后分离骨髓,采用贴壁法在体外培养、传代、扩增、纯化其骨髓间质干细胞,分别观察原代和传代后第2,4,6,8代的细胞生长情况,并绘制生长曲线和贴壁率曲线。生长曲线:以平均每孔的细胞数(千个)为纵坐标,天数(d)为横坐标;贴壁率曲线:以贴壁率(%)为纵坐标,时间(h)为横坐标。贴壁率又称接种存活率:贴壁率(%)=贴壁细胞数/接种的细胞总数×100%。结果:①骨髓细胞刚接种时呈圆型,24h后可见细胞贴壁,贴壁细胞以单核细胞为主,在72h后逐渐转变为多种形态的细胞。②大鼠骨髓间质干细胞原代细胞第1周生长速度较慢,贴壁时间长,而1周后生长加快;生长曲线及贴壁率曲线显示,传代后7代以内细胞生长速度、贴壁率基本相同;而第8代以后生长速度明显变慢,贴壁率也下降。结论:骨髓间质干细胞有一定的增殖能力,但其增殖不是无限的,为进一步了解和掌握骨髓间质干细胞的生物学特性和研究应用打下基础。AIM： To observe the feature of extraction, separation, culture and passage of mesenchymal stem cells of rat in vitro and provide experimental background for further study. METHODS： The experiment was conducted at the laboratory of neurology, Renji Hospital affiliated to Shanghai Second Medical University from June to December 2004. Youth male SD rats were selected. The bone marrow was separated after killing the rats. Bone mesenchymal stem cells with adherence were cultured, passaged, amplified and purified/n vitro. Living characteristics of the primary, 2^nd, 4^th, 6^th and 8^th generations was observed, respectively. Growth curve and the adhesive rate curve were drawn. The growth curve： ordinate-cell numbers （thousand） in every well averagely, abscissa-days （d）; The adhesive rate curve; ordinate-adhesive rate （%）, abscissa-hours （h）（adhesive rate also meant inoculation survivor rate, adhesive rate （%）=adbesive cell numbers / total inoculation cell numbers ×100%）. RESULTS： ①The marrow cells were round at the beginning and adhered in 24 hours. Most adherent cells were mainly monocytes, and changed into multiple cellular shapes gradually after 72 hours. ②The growth velocity of primary generation was slow and the adhesive time was long at the first week, but after 1 week the growth velocity was faster. The growth velocity and adhesive rate was similar before the 7^th generation. After the 8^th generation, the growth velocity was significantly dropped, and the adherence rate also decreased. CONCLUSION：Subcultured bone mesenchymal stem cells have proliferative capacity, which are limited in generations. The study provides us better background for further understanding and mastering of biological characteristics and study application of mesenchymal stem cells.

关 键 词：骨髓 间质细胞 干细胞 

分 类 号：R394.2[医药卫生—医学遗传学]