Human EPO gene eukaryotic expression vector and its application in human umbilical cord blood mesenchymal stem cells.docVIP

 PAGE \* MERGEFORMAT 16 Human EPO gene eukaryotic expression vector and its application in human umbilical cord blood mesenchymal stem cells Author: Xia Gui-zhi, ZHANG Guo-cheng, Chen Xinmin, Xin-Ru [Abstract] Objective: To construct human EPO gene eukaryotic expression vector and transfected into human umbilical cord blood mesenchymal stem cells, to obtain stable expression of human EPO in human umbilical cord blood mesenchymal stem cells. Methods: from human fetal liver cells to extract total RNA, After human EPO was amplified by RTPCR full-length cDNA, application of recombinant EPO cDNA gene was subcloned into the pcDNA3 eukaryotic expression vector in order to digestion and sequencing recombinant plasmid pcDNA3-EPO construct validity, and then pcDNA3 - EPO by the liposome-mediated transfection of human umbilical cord blood mesenchymal stem cells by G418 selection, was stably transfected EPO gene in human umbilical cord blood mesenchymal stem cells, using RT-PCR, Western Blot and cell immuno-chemical method of identification of foreign source of human EPO gene in human umbilical cord blood mesenchymal stem cells. Results: digestion and sequencing results were confirmed pcDNA3-EPO construct correct, RT-PCR, Western blot and cell immuno-chemical results showed that transfected human EPO gene in human umbilical cord blood mesenchymal stem cells in vitro to express EPO. Conclusion: The human EPO gene was successfully constructed the eukaryotic expression vector was transfected into human umbilical cord blood mesenchymal stem cells in vitro availability of stable expression. [Keywords:] rhHPO fetal blood mesenchymal stem cell transfection [Abstract] AIM: To construct the eukaryotic expression vector of human EPO gene and explore its expression in mesenchymal stem cells (MSCs) derived from human umbilical cord blood (UCB). METHODS: The total RNA was extracted from human fetal liver cells. EPO cDNA was obtained by RT-PCR amplication and subcloned into pcDNA3