胎儿骨髓间充质干细胞生物学特征及荧光标记  

The biological character and fluorescent labeling of the fetal bone mesenchymal stem cells

作　　者：田静[1] 王蓓[1] 张建华[2] 姜蓉[1] 汪维伟[1] 

机构地区：[1]重庆医科大学基础医学院组织胚胎学教研室 [2]重庆医科大学附属第一医院妇产科,重庆400016

出　　处：《重庆医科大学学报》2008年第6期665-668,672,共5页Journal of Chongqing Medical University

摘　　要：目的:探讨分离、培养、扩增和荧光标记胎儿骨髓间充质干细胞(Bore mesenchymal stem cell,BMSCs)的方法及其生物学特征。方法:取4～5个月胎儿四肢骨骨髓,Ficoll离心、贴壁筛选法分离BMSCs,体外扩增,传至第12代。检测BMSCs的生长曲线,细胞周期和细胞表型,酶活性及糖原含量,以及DAPI标记BMSCs的标记率。结果:(1)BMSCs主要呈长梭形,漩涡状排列。传代细胞生长快于原代细胞,12代后的细胞出现衰老征象;(2)流式细胞术检测第3、8代细胞表达CD29、CD71,不表达CD34、HLA-DR;G0和G1期细胞约为93.73%,S、M和G2期为6.27%;(3)第5代细胞的非特异性酯酶及糖原均为阳性,碱性磷酸酶为弱阳性;(4)DAPI标记后的BMSCs生长正常,标记率为100%,后逐渐减弱,3周时标记率仅15%。结论:分离获得的胎儿BMSCs具有MSCs的特征。DAPI标记是短期示踪BMSCs的有效方法。Objective：To explore the isolation,culture,proliferation and fluor-labelling of the fetal bone mesenchymal stem cell （BMSCs）,and their biological character. Methods：Limb bone marrow samples from fetus aged 4 to 5 months were obtained, BMSCs were isolated by Ficoll centrifugalization and adherence screening method,and then cuhurod,proliferated to the 12th generation in vitro. The growth curve,cell cycle,phenotype,enzymatic activity,glycogen and the DAPI fluor-labelling of BMSCs were observed. Results：（1）Major BMSCs were fusiform, arranged as whirlpool. After the 12^th generation,BMSCs presented the signs of senility. The subcuhured cells proliferated faster than primary cells. （2）The 3rd and 8th generation of BMSCs expressed CD29,D71,but not CD34,HLA-DR;the G0, G1 period cells were 93.73%,and the S,M,G2 period cells were 6.27%. （3）The 5th generation cells were positive of nonspecific esterase and glycogen,but weak positive of alkaline phosphatase. （4）DAPI labeled BMSCs grew normaly,and the marking rate of BMSCs was 100%,but decreased gradually,and only 15% cells were labbelled for the 3^th week. Conclusion：The acquired fetal BMSCs have the character of MSCs. The DAPI labeling is a modus operandi for the short period tracing BMSCs.

关 键 词：胎儿 骨髓间充质干细胞 分离培养 DAPI 

分 类 号：Q254[生物学—细胞生物学]