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《Acta Academiae Medicinae Militaris Tertiae》 2006-24
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Retroviral vector mediated fibronectin-thrombopoietin gene modification to human bone marrow mesenchymal stem cells

YU Jie,ZHANG Lei,MOU Shu,YANG Guang,LI Xin,XIAN Ying(Department of Hematology,Children's Hospital,Chongqing University of Medical Sciences,Chongqing 400014,China)  
Objective To modify human bone morrow mesenchymal stem cells(MSCs)with fibronectin-thrombopoietin(Fn-TPO)gene carried retroviral vector and investigate the influence of Fn-TPO gene modification on human bone marrow MSCs.Methods The retroviral vector containing Fn-TPO gene was constructed and transfected into bone marrow MSCs.The proliferation capacity of bone marrow MSCs after gene modification was assayed by trypan blue exclusion assay.The transcription of Fn-TPO gene was assayed by RT-PCR.The capacity of bone marrow MSCs to adhere hematopoietic cells was assayed by MTT and the concentration of TPO in the bone marrow MSCs culture supernatant was assayed by ELISA.Results Retroviral vector containing Fn-TPO gene was successfully constructed and modified bone morrow MSCs.Fn-TPO gene was expressed by bone morrow MSCs after gene modification.The viability of MSCs had no significant difference between pre-gene-modified and post-gene-modified(P0.05).The adhering ability of gene modified bone marrow MSCs to hematopoietic cells was reinforced(P=0.01).The concentration of TPO in the bone morrow MSCs culture supernatant rose from(5.58±0.37)ng/ml to(7.46±0.59)ng/ml(P0.01)and would not significantly decline in short time period,but was influenced by MSCs growth status.Conclusion Human bone morrow MSCs was successfully modified by Fn-TPO gene carried retroviral vector and the status of gene modified MSCs was assayed.This would facilitate further study of fibronectin and thrombopoietin genes on hematopoietic regulation and hematological system diseases.
【Fund】: 国家自然科学基金资助项目(30100095)~~
【CateGory Index】: R346
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