The Stable Expression of Vascular Endothelial Growth Factor Gene Mediated by Lipofect AMINE in Human Umbilical Vein Endothelial Cell Line Cells

Aim To establish a human umbilical vein endothelial cell (hUVEC) line monoclonal cells with the stable expression of vascular endothelial growth factor (VEGF) gene. Methods The eukaryotic expression vector PCD 2 VEGF 121 were transfected into hUVEC line cells mediated by lipofect AMINE. The positive clones were obtained by the screen of G 418 . The transcription and expression of VEGF gene were investigated by RT PCR and immunocytochemistry respectively. The experiment of Miles was applied for the assay of the biological activity of the protein of the VEGF produced by the hUVEC line cells which transfected PCD 2 VEGF 121 . The growth curve was made for comparison with that of non transfected hUVEC line cells. Results A positive clone cells from which transcripted the MRNA of VEGF 121 gene was obtained by RT PCR; The positive results of the immunocytochemistry were found and the highly biological activity of VEGF in the media was detected each only in the positive clone cells. The doubling time of the positive colon cells was shorter than that of the non transfected hUVEC line cells calculated from the growth curve. Conclusion The hUVEC line monoclonal cells with the stable expression of VEGF gene has been established successfully.