Establishment and identification of porcine aorta endothelial cell lines

Abstract

Objective To establish pig aortic endothelial cell lines for providing necessary cell model for xenotransplantation research.Methods The cell lines were established following the transfection of primary endothelial cells isolated from the aortas of the immature pig with lentivirus carrying genes for neomycin resistance and Simian Virus 40 Large T (SV40LT) antigen.The SV40LT mRNA,α-1,3-galactosyltransferase (α-1,3-Gal) mRNA and α-1,3-galactose were detected.At last,the response between the cells and human serum was tested.Results The expression of the SV40LT mRNA in cell lines were very high (P ＜0.05).Doubling time of the primary cells and the cell lines was about 21 h and 18 h,respectively.The cells also displayed endothelial cell characteristics since they endocytosed acetylated low density lipoproteins and steadily continuously expressed the yon Willebrand factor (P ＞ 0.05).The high expression of α-1,3-Gal mRNA and α-1,3-galactose [the fluorescence intensity was (498.2 ± 3.5) and (500.6 ± 3.2),respectively] had no difference in the two types of cells (P ＞ 0.05).When co-incubation with human serum,human natural antibodies bound to the two types of cells.Conclusion These results suggest that pig endothelial cell lines retained their original morphological structure and phenotypic characterization,and also had ability to respond to human serum. Key words: Pig; Endothelial cell; Xenotransplantation