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外周血基质干细胞培养鉴定及其向施万细胞的诱导分化

Isolation and identification of the peripheral blood derived mesenchymal stem cells and the induction of its into Schwann cells

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【作者】 戴翔占小多江沿文汤银娟张丽华刘忠英董为人郭家松

【Author】 DAI Xiang,ZHAN Xiao-duo,GUO Jia-song,et al. Department of Embryology and Histology,School of Basic Medical Sciences,Southern Medical University,Guangzhou 510515,China

【机构】 南方医科大学基础医学院组织学与胚胎学教研室

【摘要】 目的研究体外分离培养大鼠外周血基质干细胞(PBMSCs)并诱导分化为施万细胞的潜能。方法从SD大鼠取血分离培养PBMSCs,采用流式细胞术和免疫细胞化学法对其细胞表面抗原进行检测与鉴定,并用免疫细胞化学法检测BrdU作用4h后BrdU的阳性率。PBMSCs经β-巯基乙醇、全反式维甲酸及复合条件培养基等三个步骤向施万细胞定向诱导后,用免疫细胞化学法检测S-100和P75的表达。结果流式细胞术显示培养获得的PBMSCs中CD11b、CD29、CD45、CD49d、CD90及CD106的阳性率分别为19.97%、99.96%、46.62%、5.46%、71.22%和10.76%。免疫细胞化学法显示PBMSCs呈CD34阴性,而BrdU阳性率为(34.1±4.3)%。PBMSCs经定向诱导后S-100和P75的阳性率分别是(75.2±4.1)%和(78.9±4.6)%。结论从外周血分离获得的干细胞符合基质干细胞的特性,这些细胞在特定的条件下可诱导分化成为施万细胞。

【Abstract】 Objective To isolate and identify the peripheral blood derived mesenchymal stem cells (PBMSCs ) and the induction of it into Schwann cells in vitro. Methods: PBMSCs was isolated from SD rat blood,then special surface markers of the PBMSCs were identified by flow cytometry and immunocytochemistry. After the induction of PBMSCs towards Schwann cells successively with β-mercaptoethanol,all-trans retinoic acid and compound induce medium,the expression of S-100 or P75 was detected with immunocytochemistry. Results Flow Cytometry showed that the positive rates of CD11b,CD29,CD45,CD49d,CD90 and CD106 in cultured PBMSCs were 19.97%,99.96%,46.62%,5.46%,71.22% and 10.76% respectively. The immunocytochemistry demonstrated the negative expression of CD34 in PBMSCs. The percentage of the BrdU positive cells in PBMSCs was (34.1±4.3)% after BrdU treatment about 4h. The positive rates of S-100 and P75 were (75.2±4.1)% and (78.9±4.6)% respectively after inducing into Schwann cells. Conclusions Stem cells isolated from peripheral blood have the characteristics of mesenchymal stem cells,which can be induced into Schwann cells under specific conditions.

  • 【文献出处】 中国临床解剖学杂志 ,Chinese Journal of Clinical Anatomy , 编辑部邮箱 ,2010年02期
  • 【分类号】R329
  • 【被引频次】4
  • 【下载频次】176
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