Isolation and Growth of Endothelial Cells from the Microvessels of the Newborn Human Foreskin in Cell Culture

Abstract

A procedure for the isolation and in vitro cultivation of endothelial cells from the microvessels of the newborn human foreskin dermis is described. The epidermis was removed from foreskin tissue using a Castroviejo keratotome (0.1 mm shim). Endothelial cells were released from the dermal vessels by trypsinization of 5 mm2 sections of dermis at 37 degrees C for 40 min. Cells were expressed into Minimal Essential Medium (MEM) containing 10% pooled human serum, collected by centrifugation and plated onto either a plain plastic or a fibronectin treated culture surface. In primary culture the rate of endothelial cell proliferation was dependent upon serum type and concentration being optimal in 50% pooled human serum. High serum concentration in combination with pretreatment of the culture surface with fibronectin was required for maximal proliferation rate, for the cells to achieve confluence and for subcultivation. Primary and subcultured cells were characterized as endothelial by light microscopic, immunofluorescent (Factor VIII associated protein) and ultrastructural (Weibel-Palade body) criteria.

In vivo upregulation of CD95 and CD95L causes synergistic inhibition of angiogenesis by TSP1 peptide and metronomic doxorubicin treatment

“…HUVECs from umbilical cords isolated as described 65 or purchased from Clonetics (Cambrex BioSciences, Rockland, MD) were maintained as in Lorenzo et al 18 HMVECs isolated from neonatal foreskins [66][67][68] or purchased from Clonetics were maintained in MCDB131 medium (BioWhittaker, Walkersville, MD, USA) supplemented with 10 mM HEPES, 15% FCS, 10 ng/ml EGF (Promega, Madison, WI, USA), 2.5 mg/ml Fungizone, penicillin, streptomycin, 1 mg/ml hydrocortisone and 2 mM Lglutamine. Bladder cancer cell line 253J, B-V was maintained in DMEM, 10% FCS, 2 mM L-glutamine, penicillin and streptomycin.…”

Cutting Edge: C-C Chemokine Receptor 6 Is Essential for Arrest of a Subset of Memory T Cells on Activated Dermal Microvascular Endothelial Cells Under Physiologic Flow Conditions In Vitro

“…HDMEC were isolated from neonatal foreskins, cultured as described (17,18), and used for these studies in passages 3-6.…”

Effect of eye-derived growth factors from rabbit retina on proliferation of endothelial cell from marginal vessels.

“…As angiogenesis occurred from peripheral vessels rather than large vessels, it seems more reasonable to study the action of EDGF on the endothelial cells from the peripheral vessels than large vessels. Requirements of endothelial cells are different in their origins ; the peripheral cells require specific growth factors and treatment of Petri dishes with fibronectin or gelatin for their growth (Folkman et al 1979 ;Davison et al 1980b). We obtained the endothelial cells from the rabbit marginal vessels by the method of Davison et al (1980a) with minor modifications.…”