Basic & Clinical Medicine ›› 2014, Vol. 34 ›› Issue (6): 802-809.

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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

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