【摘要】 目的 探讨人血管内皮细胞生长因子(VECF)基因修饰的免骨髓基质细胞对VEGF基因的表达及表达产物的生物学活性。方法 制备真核表达质粒pAdCMV-VEGF165,以Superfect转染试剂介导VFGF基因转染兔骨髓基质细胞,收获转染后24h至6d的转染上清和细胞,采用RT-PCR技术、免疫组化SABC法检测转染细胞中外源性VECF165基因的转录及表达,用血管内皮细胞增殖试验测定转染细胞培养上清中VEGF的生物活性。结果 VEGF基因转染的兔骨髓基质细胞可有效转录VEGF165,其分泌于培养上清中的表达产物可促进血管内皮细胞增殖,具有很强的生物活性。结论 VEGF基因转染的兔骨髓基质细胞可有效表达具有生物活性的VEGF。更多还原

【Abstract】 Objective To explore VEGF gene expression and bioactivity of expression production by assessing VEGF gene modified rabbit bone marrow stromal cells. Methods pAdCMV - VEGF165 plasmids were extracted from DH5 a E coli and were transferred into rabbit bone marrow stromal cells mediated by Superfect Reagent. The culture media and cells were collected each 24h for 6d. The transcription and expression of VEGF gene were investigated by RT - PCR and Streptavidin - Biotin - enzyme Complex (SABC) respectively. The activity of VEGF in culture media was detected by proliferation effects on rabbit aortic vascular endothelial cells. Results The expression of VEGF gene were found distinctly in trans-fected rabbit marrow stromal cells and VEGF in the media of transfected cells had highly biological activity on proliferation of rabbit aortic vascular endothelial cells. Conclusion VEGF gene transferred rabbit marrow stromal cells could express VEGF with highly biological activity, which provided the foundation on learning VEGF gene and marrow stromal cell based gene therapy for bone repairing and regeneration.更多还原