BrdU, CFSE和GFP标记大鼠间充质干细胞的比较.doc

BrdU, CFSE和GFP标记大鼠间充质干细胞的比较
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作者:付霞霏,何援利,杨芳,彭冬先,刘木彪
【摘要】目的: 研究BrdU, CFSE和GFP作为大鼠间充质干细胞标记物的可行性. 方法: 分别用上述三种标记物标记大鼠间充质干细胞,检测即时、15和30 d的标记率,通过检测细胞周期、凋亡率和描绘生长曲线,来明确标记物对体外培养大鼠间充质干细胞生长特性的影响;通过流式细胞仪检测标记物对细胞表面标志(CD29, CD34, CD44, CD45)表达的影响. 结果: BrdU, CFSE, %, %, %,1 %, %, %. 三者对体外培养的大鼠间充质干细胞的生长特性和表面标志的表达均无明显影响. 结论: GFP, BrdU和CFSE均可以作为大鼠间充质干细胞的标记物.
【关键词】绿色荧光蛋白;溴脱氧尿嘧啶核苷;荧光染料CFSE;间充质干细胞
【Abstract】 AIM: To investigate the feasibility of labeling of BrdU, CFSE and GFP as labels of rat mesenchymal stem cells (MSCs) and to investigate the influence of the 3 markers on the growth characteristics and expressions of surface markers of rat MSCs. METHODS: Wistar rats
 bone marrow derived MSCs were isolated and purified in vitro by Percoll density gradient bined with adherent method and were labeled with BrdU, CFSE and GFP, respectively. The immediate, 15day and 30day labeling efficiency of the 3 markers was quantified. The labeling efficiency of BrdU was detected with immunocytochemical method, and that of CFSE and GFP was detected via flow cytometry (FCM). The labeled MSCs were observed under microscope. Cell cycle and apoptosis rate of labeled MSCs were revealed and the growth curves were delineated, so as to identify whether or not the markers casted an influence on the cell growth characteristics in vitro. Furthermore, the surface markers (CD29, CD34, CD44, CD45) of the labeled MSCs were detected through flow cytometry and pared with those of nonlabeled MSCs. RESULTS: The immediate labeling efficiency of BrdU, CFSE, and GFP was %, %, % respectively, and the efficiency decreased to %, %, and % respectively after 30 d. There was no statistical difference in cell cycle and ap
optosis ratio between labeled MSCs and nonlabeled MSCs. The growth curves of labeled MSCs were similar to that of nonlabeled MSCs. The 3 markers didnt influence the expressions of surf