血管内皮细胞生长因子基因转染皮肤成纤维细胞的实验研究

为改善人工复合皮在临床上的应用效果 ,构建人血管内皮细胞生长因子 16 5 (hVEGF165)基因重组载体pcDNA3,并转染成纤维细胞。检测转基因细胞培养上清液VEGF浓度 ;用转基因细胞培养上清液刺激人脐静脉血管内皮细胞 ,观察内皮细胞增殖速度 ;通过Miles实验检测VEGF的生物学活性。结果显示 ,转基因成纤维细胞能够表达一定浓度的VEGF,且具有加速体外培养人脐静脉血管内皮细胞 (HUVEC)生长和增强血管通透性的生物学活性。提示转hVEGF165基因成纤维细胞可修饰人工皮的真皮面。

EXPERTMENTAL STUDIES ON SKIN FIBROBLASTS TRANSFECTED WITH HUMAN VASCULAR ENDOTHELIAL GROWTH FACTOR GENE

This study was aimed to improve the therapeutic results of composite skin substitutes. Human vascular endothelial cell growth factor (hVEGF 165 ) gene was constructed to an eukaryotic expression vector pcDNA3. Transfection of recombinant vector pcDNA3 hVEGF 165 into human dermal fibroblast cells was performed. VEGF protein level in the supernatant of transfected fibroblasts culture was determined. Its biological activities were tested by observing the growth rate of the human umbilical vein endothelial cells/HUVEC after being stimulated with the said supernatant, and by performing the Miles assay in guinea pigs. The results showed that these transgenic cells were able to secrete VEGF to certain extent, with biological activities to enhance the growth of HUVEC in vitro and improve vascular permeability. It indicated that transgenic fibroblasts could resurface the dermal substitute of a composite skin.