Brain- and liver cell-derived factors are required for growth of human endothelial cells in serum-free culture.

Abstract

A test of the mitogenic activity of >40 purified and crude sources of hormones and growth factors revealed that epidermal growth factor, high density lipoprotein, an extract of bovine pituitary, hypothalamus, or whole brain, and the medium conditioned by differentiated human hepatoma cells were (9,10), and an acid pituitary-or brain-derived factor (5,11,12) are active mitogens for endothelial cells derived from human umbilical vein (HUVE). In addition, thrombin has been reported to potentiate the action of EGF, basic fibroblast growth factor (6-8), and partially purified platelet-derived growth factor (9). Although there is no consensus on the importance of platelet mitogens (5), thrombin (7), and the basic (5, 11) and acidic (8) growth factors from brain, it appears agreed that EGF and one or more factors derived from neural tissue are mitogens for HUVE (5-8). However, in the presence of both EGF and brain-derived factors, HUVE stillThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact. 6413have a fastidious requirement for unidentified plasma-derived factors (5-12).Our studies show that, in addition to EGF and brain-derived factors, one or more factors from the medium of differentiated human hepatoma cells is required for HUVE proliferation in the absence of serum. This provides a serum-free medium in which to study and clarify the endocrinology of growth, function, and aging of cultured human endothelial cells.

Interaction of Leptospira interrogans with Human Proteolytic Systems Enhances Dissemination through Endothelial Cells and Protease Levels

“…The HUVECs were obtained by collagenase digestion of umbilical veins as detailed by Jaffe et al (18). The cells were cultured in RPMI 1640 medium (Cultilab) supplemented with 10% fetal bovine sera (FBS) (Cultilab), 45 g/ml heparin (Roche), 1 mM sodium pyruvate (Spectrum Chemical), 2 mM L-glutamine (Sigma), 100 IU penicillin (Cultilab), 100 g/ml streptomycin (Cultilab), 50 M 2-mercaptoethanol (Pharmacia Biotech), 25 g/ml endothelial growth factor (Sigma), and 0.75% mouse brain extract, as a supplement of growth factor (19). The latter was prepared by homogenization of approximately 5 g of mouse brain in 5 ml of RPMI medium, as described by Maciag et al (20).…”

Endothelial dysfunction in a rat model of endotoxic shock. Importance of the activation of poly (ADP-ribose) synthetase by peroxynitrite.

“…Human umbilical vein endothelial cells (HUVEC; American Type Culture Collection, Rockville, MD) were cultured in F12K medium, 10% FBS, 100 g/ml heparin, and 30 g/ml endothelial cell growth supplement as described previously (17). Mouse embyo fibroblasts from the PARS Ϫ / Ϫ mouse and fibroblasts from the corresponding wild-type controls (18) were grown in DME with 10% FBS.…”

Role of lipoproteins in growth of human adult arterial endothelial and smooth muscle cells in low lipoprotein‐deficient serum

“…The SMC were isolated from the deendothelialized tissue according to the procedures described for EC isolation except that SMC were scraped from collagenasedigested tissue with a scalpel instead of a cotton swab. Cells (EC and SMC) were seeded in collagen-coated T-25 tissue culture flasks (Hoshi and McKeehan, 1984) in 5 ml of medium MCDB 107 supplemented with EGF (10 ng/ml), partially purified bovine brain-derived growth factor(s) (Gordon et al, 19831, and 2% fetal bovine serum (FBS). For the growth of EC, the medium was further supplemented with 100 pg/ml of heparin.…”