|Expression of human VEGF(121) cDNA in mouse bone marrow stromal cells|
|Cai, SB; Ma, QJ; Yu, XJ; Dang, GT; Ma, DL|
|关键词||endothelial growth factor retrovirus vector gene therapy marrow stromal cell|
|刊名||CHINESE MEDICAL JOURNAL|
|WOS标题词||Science & Technology|
|类目[WOS]||Medicine, General & Internal|
|研究领域[WOS]||General & Internal Medicine|
|关键词[WOS]||VASCULAR-PERMEABILITY FACTOR ; ENDOTHELIAL GROWTH-FACTOR ; LIMB ISCHEMIA ; ANGIOGENESIS ; GENE ; VEGF|
Objective To construct a retroviral vector carrying human vascular endothelial growth factor ( hVEGF(121) cDNA for evaluation of the possibility of VEGF gene therapy in ischemic bone disease.
Methods hVEGF(121) cDNA was obtained from the plasmid pCDI/VEGF(121), and cloned into retroviral plasmid pLXSN. Recombinant plasmid was transferred to the retrovirus packaging cell, PT-67, by lipofectamine mediated gene transfer. Mouse bone marrow stromal cells (MSCs) were transfected by the retrovirus. The integration of the hVEGF(121) cDNA into MSC genomic DNA and expression of the VEGF gene was detected. Proliferation assays of human umbilical vein endothelial cells (HUVECs) by VEGF(121) in culture medium were performed.
Results Recombinant pLXSN/VEGF(121) was correctly constructed and confirmed by restriction endonuclease analysis and DNA sequencing analysis. hVEGF(121) gene was integrated into MSC genomic DNA after transfection, and the VEGF(121) protein was expressed. Proliferation assays showed VEGF(121) in culture medium was a biologically active protein and had a mitogenic effect on HUVEC.
Conclusions Recombinant retroviral vector carrying hVEGF(121) cDNA was successfully constructed. VEGF(121) protein expressed by MSCs had mitogenic effect biologically. This provides a further foundation for VEGF gene therapy for bone ischemic disease and bone tissue engineering.
|作者单位||1.Peking Univ, Dept Orthoped, Hosp 3, Beijing 100083, Peoples R China|
2.Peking Univ, Ctr Human Dis Genom, Beijing 100083, Peoples R China
|Cai, SB,Ma, QJ,Yu, XJ,et al. Expression of human VEGF(121) cDNA in mouse bone marrow stromal cells[J]. CHINESE MEDICAL JOURNAL,2002,115(6):914-918.|
|APA||Cai, SB,Ma, QJ,Yu, XJ,Dang, GT,&Ma, DL.(2002).Expression of human VEGF(121) cDNA in mouse bone marrow stromal cells.CHINESE MEDICAL JOURNAL,115(6),914-918.|
|MLA||Cai, SB,et al."Expression of human VEGF(121) cDNA in mouse bone marrow stromal cells".CHINESE MEDICAL JOURNAL 115.6(2002):914-918.|