title: Establishment and characterization of endothelial cell lines from the aorta of miniature pig for the study of xenotransplantation.
abstract: Pig endothelial cells are the first cells to interact with human immune components after organ xenotransplantation, which is a procedure currently considered to be the best treatment option for end-stage organ failure. It is, therefore, essential to study the mechanisms of molecular interaction between pig endothelial cells and human immune components, in order to overcome xenograft rejection. The aim of this study was to establish immortalized pig aortic endothelial cell lines, in order to facilitate future in vitro studies of human anti-pig immune responses. Endothelial cell lines were established following the transfection of primary endothelial cells isolated from the aortas of the Minnesota miniature pig with plasmid pRNS-1 carrying genes for neomycin resistance and the SV40 large T antigen. The immortalized cell lines showed a relatively rapid doubling time (17.6h) and the endothelial cell phenotype, as indicated by the formation of typical cobblestone monolayers and by the constitutive expression of PECAM-1 and the von Willebrand factor. Flow cytometric analysis demonstrated the constitutive expression of SLA class I and CD86, whereas the expression of E-selectin and SLA class II was only induced after stimulation with human TNF-alpha and pig IFN-gamma, respectively. On the other hand, no CD80 expression was detected in the primary cells or cell lines in the presence or absence of either human TNF-alpha or pig IFN-gamma. A vigorous human T cell proliferation against these cell lines was observed in the mixed lymphocyte-endothelial cell culture. These results suggest that pig endothelial cells, immortalized by the introduction of SV40 T, retain their original characteristics, except for the acquired property of immortalization, and that they may be useful for future in vitro studies of xenogeneic human anti-pig immune responses.
authors.affiliation: Xenotransplantation Research Center and Transplantation Research Institute, Seoul National University Hospital, Republic of Korea.
authors.forename: Donghee
authors.initials: D
authors.lastname: Kim
authors.forename: Jae Young
authors.initials: JY
authors.lastname: Kim
authors.forename: Hyun Sook
authors.initials: HS
authors.lastname: Koh
authors.forename: Jung Pyo
authors.initials: JP
authors.lastname: Lee
authors.forename: Young Tae
authors.initials: YT
authors.lastname: Kim
authors.forename: Hee Jung
authors.initials: HJ
authors.lastname: Kang
authors.forename: Woo Suk
authors.initials: WS
authors.lastname: Hwang
authors.forename: Yoon Berm
authors.initials: YB
authors.lastname: Kim
authors.forename: Jung Sang
authors.initials: JS
authors.lastname: Lee
authors.forename: Curie
authors.initials: C
authors.lastname: Ahn
mesh_terms: D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics*; D001011:Aorta / Q000166:cytology*; D002461:Cell Line, Transformed; D049109:Cell Proliferation; D002472:Cell Transformation, Viral; D046508:Culture Techniques; D019040:E-Selectin; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D013552:Swine; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D014183:Transplantation, Heterologous; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014841:von Willebrand Factor / Q000378:metabolism
chemical_list: D000952:Antigens, Polyomavirus Transforming; D019040:E-Selectin; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D014409:Tumor Necrosis Factor-alpha; D014841:von Willebrand Factor; D007371:Interferon-gamma