中国畜牧兽医

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绵羊羊水、胎儿肾脏组织中表达Oct-4细胞的分离培养及其分子学特性研究

王凤武1，2，郝斐3，袁建龙3，刘东军3，荣威恒2   

  1. (1.内蒙古农业大学动物科学学院，内蒙古呼和浩特  010018；2.内蒙古农牧业科学院，内蒙古呼和浩特  010031；3.内蒙古大学国家动物转基因技术中心，内蒙古呼和浩特  010021)
  • 出版日期:2014-01-20 发布日期:2014-02-27
  • 作者简介:王凤武（1973—），男，内蒙古人，博士，研究方向：分子生物学与牛羊育种。
  • 基金资助:

    内蒙古自然科学基金（2011MS0410）。

Comparative Analysis in Molecular Characteristic of the Oct-4 expressing Cells Isolated from Sheep Amniotic Fluid and Fetal Renal Tissues

WANG Feng-wu1,2, HAO Fei3, YUAN Jian-long3， LIU Dong-jun3, RONG Wei-heng2   

  1. (1.College of Animal Science,Inner Mongolia Agricultural University,Hohhot 010018,China； 2.Inner Mongolia Academy of Agricultural and Animal Husbandry, Hohhot 010031,China； 3.The National Animal Transgenic Technology Centre, Inner Mongolia University,Hohhot 010021,China)
  • Online:2014-01-20 Published:2014-02-27

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摘要： 试验旨在建立从绵羊羊水、胎儿肾脏组织中分离培养表达Oct-4细胞的新方法，同时，通过实时荧光定量PCR（Real-time PCR）初步探索二者在分子生物学特性方面的联系及差异。采用0.01%胰酶培养液，从同一只受孕中期绵羊的羊水和胎儿肾脏组织中各分离1株细胞。Real-time PCR分析结果表明，从绵羊羊水、胎儿肾脏组织中分离的细胞均表达胚胎干细胞相关标志基因Oct-4、胚胎生殖细胞标志基因SSEA-1、主要组织相容性抗原MHC-Ⅱ、细胞凋亡基因Bax及抑制细胞凋亡基因Bcl-2，不表达MHC-Ⅰ、Nanog及TERT，因此，将羊水中表达Oct-4的细胞暂命名为羊水来源多潜能干细胞(amniotic fluid stem cells，AFSC),胎儿肾脏组织中表达Oct-4的细胞暂命名为肾脏组织干细胞(renal tissue stem cells，RTSC)。相对定量分析结果显示，二者的Oct-4、Bax及Bcl-2基因的相对表达量存在明显差异。绵羊羊水中表达Oct-4的细胞（AFSC）和胎儿肾脏组织中表达Oct-4的细胞（RTSC）体外悬浮培养7 d均能形成类胚体。试验成功建立了从绵羊羊水、胎儿肾脏组织中分离培养表达Oct-4细胞的新方法，实时荧光定量PCR分析初步显示绵羊羊水中表达Oct-4的细胞和胎儿肾脏组织中表达Oct-4的细胞具有相同的起源，并在体内处于一种动态的变化之中。

关键词: 绵羊; 0.01%胰酶培养液; 羊水; 胎儿肾脏组织; Oct-4; 细胞; 类胚体

Abstract: This study aimed at developing a novel technique of isolating the Oct-4 expressing cells from the sheep amniotic fluid and fetal renal tissues, respectively, and comparatively analyzing the difference in the molecular characteristic of these two differently derived cells with Real-time PCR. The results showed that the cells from the sheep amniotic fluid and fetal renal tissue were both positive for Oct-4, embryonic germ cells gene SSEA-1, the main organization compatibility antigen gene MHC-Ⅱ, apoptosis gene Bax, and inhibition gene of apoptosis Bcl-2, negative for the symbol marker of MHC-Ⅰ, Nanog and TERT, but there were marked differences in the relative expression levels of Oct-4， Bax and Bcl-2 genes. Furthermore, AFSC and RTSC were both capable of forming embryoid bodies through the suspension culture in vitro. In conclusion, our study successfully established a new procedure of isolating and culturing Oct-4 expressing cells derived from the sheep amniotic fluid and fetal renal tissues, which might have the same origin.

Key words: sheep; 0.01% trypsin medium; sheep amniotic fluid; fetal renal tissue; Oct-4; cell; embryoid bodies

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