Expression of human VEGF(121) cDNA in mouse bone marrow stromal cells / 中华医学杂志(英文版)
Chinese Medical Journal
;
(24): 914-918, 2002.
Artigo
em Inglês
| WPRIM
| ID: wpr-302275
ABSTRACT
<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>hVEGF(121) cDNA was obtained from the plasmid pCDI/VEGF(121) and cloned into retroviral plasmid pLXSN. Recombinant plasmid was transferred to the retro virus 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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>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.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Plasmídeos
/
Retroviridae
/
Células da Medula Óssea
/
Endotélio Vascular
/
Terapia Genética
/
Divisão Celular
/
Fatores de Crescimento Endotelial
/
Linfocinas
/
Células Estromais
/
DNA Complementar
Limite:
Animais
/
Humanos
Idioma:
Inglês
Revista:
Chinese Medical Journal
Ano de publicação:
2002
Tipo de documento:
Artigo
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