中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2013, Vol. 8 ›› Issue (13): 1210-1219.doi: 10.3969/j.issn.1673-5374.2013.13.006

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

绵羊羊膜上皮细胞干细胞特性及向神经分化的潜能

  

  • 收稿日期:2012-12-19 修回日期:2013-04-13 出版日期:2013-05-05 发布日期:2013-05-05

Stem cell properties and neural differentiation of sheep amniotic epithelial cells

Xuemin Zhu1, 2, Xiumei Wang1, Guifang Cao1, Fengjun Liu2, Yinfeng Yang1, Xiaonan Li3, Yuling Zhang2, Yan Mi1, Junping Liu1, Lingli Zhang1   

  1. 1 Veterinary Medicine College of Inner Mongolia Agricultural University, Hohhot 010018, Inner Mongolia Autonomous Region, China
    2 College of Science and Technology, Henan University of Science and Technology, Luoyang 471023, Henan Province, China
    3 Liaoning Center for Animal Epidemic Disease Control and Prevention, Shenyang 110164, Liaoning Province, China
  • Received:2012-12-19 Revised:2013-04-13 Online:2013-05-05 Published:2013-05-05
  • Contact: Guifang Cao, M.D., Professor, Veterinary Medicine College of Inner Mongolia Agricultural University, Hohhot 010018, Inner Mongolia Autonomous Region, China, guifangcao@126.com.
  • About author:Xuemin Zhu☆, M.D., Lecturer. Xiumei Wang, Xiaonan Li and Xuemin Zhu contributed equally to this paper.

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摘要：

为了证实绵羊羊膜上皮细胞的干细胞特性和神经分化能力，实验从分娩母羊的羊膜分离培养羊膜上皮细胞，利用细胞免疫荧光和反转录PCR方法检测，发现羊膜上皮细胞表达胚胎干细胞标志蛋白SSEA-1，SSEA-3，SSEA-4，TRA-1-60和TRA-1-81和全能性相关基因Oct-4，Sox-2和 Rex-1 mRNA，不表达Nanog mRNA。通过无血清神经诱导培养液Neurobasal-A medium添加B-27的神经诱导介质诱导培养羊膜上皮细胞，在诱导后28d，细胞表达β-III-微管蛋白、神经胶原纤维酸性蛋白和标志基因巢蛋白及微管相关蛋白2等。作者认为，获得的具有干细胞特性的绵羊羊膜上皮细胞，在特定条件下能够分化为β-III-微管蛋白和微管相关蛋白2表达的具有1个轴突和2个树突或是多树突及其他形态的神经元和神经胶原纤维酸性蛋白阳性表达的神经胶质样细胞。

关键词: 神经再生, 干细胞, 绵羊, 羊膜上皮细胞, 分离培养, 干细胞特性, 分化, 神经分化潜能, 基金资助文章

Abstract:

This study was designed to verify the stem cell properties of sheep amniotic epithelial cells and their capacity for neural differentiation. Immunofluorescence microscopy and reverse transcription-PCR revealed that the sheep amniotic epithelial cells were positive for the embryonic stem cell marker proteins SSEA-1, SSEA-3, SSEA-4, TRA-1-60 and TRA-1-81, and the totipotency-associated genes Oct-4, Sox-2 and Rex-1, but negative for Nanog. Amniotic epithelial cells expressed β-III-tubulin, glial fibrillary acidic protein, nestin and microtubule-associated protein-2 at 28 days after induction with serum-free neurobasal-A medium containing B-27. Thus, sheep amniotic epithelial cells could differentiate into neurons expressing β-III-tubulin and microtubule-associated protein-2, and glial-like cells expressing glial fibrillary acidic protein, under specific conditions.