中文摘要：

许多研究人员学习了的背景把干细胞用作基因的可能性治疗学的向量。但是脂肪质的导出织物的干细胞(ADSC ) 上的很少相关报告是可得到的。因此，我们打算构造 lentiviral VEGF165 表示向量然后感染 ADSC 生产治疗学的种子 cells.Methods EHS1001-68950485313912 克隆被 PCR 方法变异生产 VEGF165 抄本(NM_001025368 ) 的一致碎片。Lentivirus 与 pGC-FU 被包， pHelper 1.0 并且 pHelper 在当时， ADSC (infection=20 的复合) 是的 293T cells.And 的 2.0 plasmids 有在 titer 决心以后的向量的 transfected。在 ADSC 的 VEGF165 的稳定的表示被 immunofluorescence 染色，连接酶的 immunosorbent 试金(ELISA ) 和西方的弄污 analysis.Results DNA 定序证实， 293T transfection 证实 VEGF165 被连接到 GFP 熔化向量。病毒 titer 被量的 PCR 决定直到 2x10a。VEGF165 transduced 房间能证明绿荧光由染色的 immunofluorescence (几乎 95 ) 证实了。ELISA 分析能外面检测 VEGF 的密度是 850.86-1202.13pg/ml (平均数(923.00 獴愠汬睯? 桴 ? 牧睯桴漠 ? 牵敯楰桴汥畩 ? 湡 ? 慣? 敳癲? 獡愠挠牡楲牥映牯琠敨琠獩?? 湥楧敮牥湩 ? 景甠敲桴慲

英文摘要：

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.