首页|期刊导航|中华医学杂志（英文版）|Over-expression of VEGF165 in the adipose tissue-derived stem cells via the lentiviral vector
中华医学杂志（英文版）2011，Vol.125Issue(19)：3093-3097,5.DOI:10.3760/cma.j.issn.0366-6999.2011.19.026
Over-expression of VEGF165 in the adipose tissue-derived stem cells via the lentiviral vector
Over-expression of VEGF165 in the adipose tissue-derived stem cells via the lentiviral vector
摘要
Abstract
Background Many researchers studied the possibility of using stem cells as gene therapeutic vector. But few related reports on the adipose tissue-derived stem cells (ADSCs) are available. Therefore we intended to construct a lentiviral VEGF165 expression vector and then infect the ADSCs to produce therapeutic seed cells.Methods EHS1001-68950485313912 clone was mutated by PCR method to produce consensus fragment of VEGF165 transcript (NM_001025368). Lentivirus was enveloped with pGC-FU, pHelper 1.0 and pHelper 2.0 plasmids in 293T cells.And then the ADSCs (multiplicity of infection=20) were transfected with the vectors after titer determination. Stable expression of VEGF165 in ADSCs was confirmed by immunofluorescence staining, enzyme-linked immunosorbent assay (ELISA) and Western blotting analysis.Results DNA sequencing and 293T transfection verified VEGF165 was linked to the GFP fused vector. The virus titer is up to 2x10a determined by quantitative PCR. VEGF165 transduced cells could show green fluorescence confirmed by immunofluorescence staining (almost 95％). ELISA analyses could detect out the density of VEGF was 850.86-1202.13pg/ml (mean (923.00±31.22) pg/ml) in the supernatant of VEGF16s-transduced cells but not detected in the GFP-transduced cells (P ＜0.001) and the Western blotting analyses also confirmed VEGF165 expression in VEGF165-transduced cells.Conclusions The VEGF165 over-expression ADSCs were obtained and may be used as a cell therapeutic tool and may be applied for vascular regeneration, especially in the treatment of erectile dysfunction.关键词
adipose tissue-derived stem cells/VEGF165/lentivirus/over-expressionKey words
adipose tissue-derived stem cells/VEGF165/lentivirus/over-expression引用本文复制引用
SUN Xiang-zhou,LIU Gui-hua,WANG Zhuo-qing,ZHENG Fu-fu,BIAN Jun,HUANG Yan-ping,GAO Yong,ZHANG Ya-dong,DENG Chun-hua..Over-expression of VEGF165 in the adipose tissue-derived stem cells via the lentiviral vector[J].中华医学杂志（英文版）,2011,125(19):3093-3097,5.基金项目
This work was supported by grants from the National Natural Science Fundation of China (No.30872571),National Youth Natural Science Fundation of China (No.30901487),Natural.Science Fund Program of Guangdong Province (No.8451008901000774), Natural Science Fund Program of Guangdong Province (No.07001628),Guangdong Province Science & Technology Project (No.2008GBCBC01250-014-149). （No.30872571）
