中国临床解剖学杂志 ›› 2010, Vol. 28 ›› Issue (2): 203-.

• 实验研究 • 上一篇    下一篇

外周血基质干细胞培养鉴定及其向施万细胞的诱导分化

戴 翔, 占小多, 江沿文, 汤银娟, 张丽华, 刘忠英, 董为人, 郭家松   

  1. 南方医科大学基础医学院组织学与胚胎学教研室,  广州   510515
  • 收稿日期:2009-12-10 出版日期:2010-03-25 发布日期:2010-04-13
  • 通讯作者: 郭家松, 副教授,硕士生导师,Tel:(020)61648204,E-mail: jiasongguo@yahoo.com.cn E-mail:ddaixiang@163.com
  • 作者简介:戴 翔(1982-),男,湖南人,硕士研究生,主要从事干细胞与脊髓损伤的研究

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

DAI Xiang, ZHAN Xiao-duo, GUO Jia-song, et al.   

  1. Department of Embryology and Histology, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China
  • Received:2009-12-10 Online:2010-03-25 Published:2010-04-13

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。

关键词: 外周血, 基质干细胞, 施万细胞, 分化

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.

Key words: Peripheral blood, Mesenchymal stem cells, Schwann cells, D ifferentiation

中图分类号: