基础医学与临床 ›› 2014, Vol. 34 ›› Issue (6): 802-809.

• 研究论文 • 上一篇    下一篇

应用组织特异性分子的保守序列鉴定人源细胞的组织来源

孙昊1,刘玉琴2,卞晓翠3,杨振丽1,赵晓梅2   

  1. 1. 中国医学科学院基础医学研究所 北京协和医学院基础学院
    2. 中国医学科学院基础医学研究所;北京协和医学院基础学院
    3. 中国医学科学院 北京协和医学院 基础医学研究所
  • 收稿日期:2014-03-19 修回日期:2014-04-11 出版日期:2014-06-05 发布日期:2014-05-26
  • 通讯作者: 刘玉琴 E-mail:ccc@pumc.edu.cn

Application of mRNA conserved sequence from Tissue-specific factors to the identification of the tissue origin of cultured cells

  • Received:2014-03-19 Revised:2014-04-11 Online:2014-06-05 Published:2014-05-26
  • Contact: Yu-qin LIU, E-mail:ccc@pumc.edu.cn

摘要: 目的 建立一种有效简便经济的检测方法,用于鉴定培养细胞的组织来源,对新鲜取材细胞的归类,检测细胞组织来源。方法 根据文献报道与NCBI数据库设计了17对组织特异性相关引物,分别针对16个常见的人类组织特异性因子的mRNA保守序列,培养细胞,提取mRNA,反转录为cDNA,以此为模板进行PCR,琼脂糖凝胶电泳分析;对这些引物进行特异性和敏感性的筛选;其中8对引物可用于判断人类组织特异性,以双盲实验对此方法进行验证,用于鉴定人类细胞的组织来源。结果 17对组织特异性相关引物中,特异性分子ALB、AFP、SYN、CDH16、SFTPB、LCA、FLT和MUL2的对应引物可有效应用于鉴定细胞是否源自肝、肾、神经、肺、造血、内皮和分泌组织;特异性因子PSA对应引物可广泛在人类各组织细胞中扩增出条带;鉴定其他组织的引物有待进一步研究完善优化。结论 应用反转录聚合酶链式反应扩增编码组织特异性分子mRNA的保守序列可以准确的鉴定待检测细胞的组织来源。本研究为鉴定常见细胞的组织来源,提供新方法。

关键词: 组织特异因子, 细胞, 组织起源

Abstract: Objective To establish an effective and economic fast method that could identify the tissue origin of cultured cells , classify fresh derived human tissue cells and different tissue cells. Methods According to references and NCBI database, we designed 17 pairs of tissue-specific primers targeted to the conserved sequences of mRNA coding human tissue-specific factors.The mRNA isolated from the known cells was reverse transcribed to cDNA which was further amplified with these primers by PCR. Agarose gel analysis of the PCR products was employed to testify specificity and sensitivity.And the selected primers were applied to identify double-blinded unknown samples. Results Among the 17 pairs of tissue-specific primers, 8 pairs corresponding to ALB, AFP, SYN, CDH16, SFTPB, LCA, FLT, MUL2 could identify whether the cell originates from the liver, kidney, nerve, lung, hematopoietic or endothelial tissue. The prostate specific antigen (PSA) can be amplified from cells of all tissue origin. Conclusion This RT-PCR assay of tissue-specific mRNA conserved fragments provides a simple ,rapid, sensitive ,and cost-effective new method to identify cell tissue types and detect cell origin.

Key words: tissue-specific factors, cell, tissue origin