caa a22 4500 477080952 CHVBK 20180405111449.0 cr unu---uuuuu 170330e19960101xx s 000 0 eng 10.1007/BF02722989 doi (NATIONALLICENCE)springer-10.1007/BF02722989 Establishment of permanent human endothelial cells achieved by transfection with SV40 large T antigen that retain typical phenotypical and functional characteristics [Elektronische Daten] [Florina Moldovan, Houda Benanni, Jean Fiet, Olivier Cussenot, Jacques Dumas, Christian Darbord, Hany Soliman] Summary: The plasmid pMK16 containing-SV40 replicated origin defective gene was efficiently introduced into early-passage human umbilical vein endothelial cells (HUVEC) using positively charged liposomes. The resulting cell line acquired an almost infinite lifespan, was morphologically unchanged, expressed SV40-antigen, and coexpressed von Willebrand factor (vWF), tissue plasminogen activator (t-PA), plasminogen activator inhibitor-1 (PAI-1), angiotensin conversion enzyme (ACE), and endothelin converting enzyme (ECE). In addition, these are the first immortalized human endothelial cells, to our knowledge, that biosynthesized and secreted interleukins (IL-1β and IL-6) in both a constitutive and regulated fashion and endothelin-1 (ET-1), the most potent vasoactive peptide, which has been suggested to be implicated in the pathogenesis of hypertension. Interestingly enough, both of the immortalized cells and the early-passage HUVEC from which the immortalized cells were obtained biosynthesized and secreted the same levels of ET-1 suggesting full maintenance of its biosynthetic pathway including the presence of active ECE, which cleaves big endothelin-1 (big-ET-1) to ET-1 and regulation factors. Moreover, the immortalized cells retained the ability to express the functional specific amino acid Na+-independent system Y+ transporter, which mediates L-arginine transport into endothelial cells from which endothelium-derived relaxing factor (EDRF, nitric oxide) is formed via the action of nitric oxide-synthase. Obtaining these immortalized human endothelial cells without alteration of the differentiated characteristics constitutes a useful model: (a) to study ET-1 secretion, gene regulation, and human ECE, which may be an important therapeutic target in disease conditions in which ET-1 is to be implicated; (b) to study L-arginine transport, which is a key step in the formation of EDRF; (c) to study IL-1β and IL-6 secretions, and gene regulations; (d) to substitute large quantities of HUVEC; and, finally, (e) to reproduce, starting with different primary endothelial cells both from human and animal origin. Society for In Vitro Biology, 1996 immortalization nationallicence endothelial cells nationallicence endothelin nationallicence interleukins nationallicence arginine nationallicence SV40 nationallicence Moldovan Florina Department of Hormonal Biology, Saint Louis University Hospital, 1, C. Vellefaux Avenue, 75010, Paris, France aut Benanni Houda Department of Microbiology Faculty of Biological Sciences, Paris V University, France aut Fiet Jean Department of Hormonal Biology, Saint Louis University Hospital, 1, C. Vellefaux Avenue, 75010, Paris, France aut Cussenot Olivier Department of Urology, Saint Louis University Hospital, 1, C. Vellefaux Avenue, 75010, Paris, France aut Dumas Jacques Biotechnology Department Roussel-UCLAF Laboratories Paris V University, France aut Darbord Christian Department of Microbiology Faculty of Biological Sciences, Paris V University, France aut Soliman Hany Department of Hormonal Biology, Saint Louis University Hospital, 1, C. Vellefaux Avenue, 75010, Paris, France aut In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 32/1(1996-01-01), 16-23 1071-2690 32:1<16 1996 32 11626 https://doi.org/10.1007/BF02722989 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF02722989 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Moldovan Florina Department of Hormonal Biology, Saint Louis University Hospital, 1, C. Vellefaux Avenue, 75010, Paris, France aut NATIONALLICENCE 700 1- Benanni Houda Department of Microbiology Faculty of Biological Sciences, Paris V University, France aut NATIONALLICENCE 700 1- Fiet Jean Department of Hormonal Biology, Saint Louis University Hospital, 1, C. Vellefaux Avenue, 75010, Paris, France aut NATIONALLICENCE 700 1- Cussenot Olivier Department of Urology, Saint Louis University Hospital, 1, C. Vellefaux Avenue, 75010, Paris, France aut NATIONALLICENCE 700 1- Dumas Jacques Biotechnology Department Roussel-UCLAF Laboratories Paris V University, France aut NATIONALLICENCE 700 1- Darbord Christian Department of Microbiology Faculty of Biological Sciences, Paris V University, France aut NATIONALLICENCE 700 1- Soliman Hany Department of Hormonal Biology, Saint Louis University Hospital, 1, C. Vellefaux Avenue, 75010, Paris, France aut NATIONALLICENCE 773 0- In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 32/1(1996-01-01), 16-23 1071-2690 32:1<16 1996 32 11626 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer