Establishment of vascular endothelial cell lines in a serum-free culture and the discovery of endothelin and a vasoactive intestinal contractor (VIC)

• Published in: Cytotechnology (Dordrecht) 1991, Vol.5 (S2), p.9-16
• Main Authors: MITSUI, Y, SAIDA, K, ISHIDA, N, IMAMURA, T, KOBAYASHI, M
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animal cells; Animals; Biological and medical sciences; Biotechnology; Cell Line; Cloning, Molecular; Culture Media; Endothelins - genetics; Endothelins - isolation & purification; Endothelins - metabolism; Endothelium, Vascular - cytology; Establishment of new cell lines, improvement of cultural methods, mass cultures; Eukaryotic cell cultures; Fundamental and applied biological sciences. Psychology; Humans; Methods. Procedures. Technologies; Peptides - genetics; Peptides - isolation & purification; Peptides - metabolism; Sequence Homology, Nucleic Acid
• ISSN: 0920-9069; 1573-0778
• DOI: 10.1007/BF00573877

Immortal vascular endothelial cell lines were established and utilized for the production of an endothelium-derived contraction factor (EDCF) in a serum-free medium. After the discovery of Endothelin (21 amino acid peptide, ET) as an EDCF, a prepro ET cDNA isolated from human tissue was used to examine the expression of ET and its regulation in human endothelial cells. A gene family of ET was shown in mouse by using prepro ET cDNA as a probe. Thus, a novel peptide, Vasoactive Intestinal Contractor (VIC) homologous to ET was deduced from the sequence of one of these genes. VIC was confirmed to induce vasocontraction as well as intestinal contraction. Northern blot analysis indicated that this gene was expressed in the intestine but not in endothelial cells. A cloning and sequencing of prepro VIC cDNA from mouse intestine suggest that a VIC-like peptide, as well as VIC, are co-synthesized by cleavage from prepro VIC with 160 amino acids.