Models of in vitro angiogenesis: endothelial cell differentiation on fibrin but not matrigel is transcriptionally dependent.
Endothelial cells can be induced to form a branching network of tubular structures using a variety of cell culture conditions. We have examined this differentiation process using several sets of conditions: plating human umbilical vein endothelial cells (HUVEC) on Matrigel, adding collagen to the apical surface of HUVEC grown on fibronectin, and plating HUVEC on fibrin in the presence of FGF-1. We determined that although the first two conditions produce dramatic morphologic changes in the HUVEC population, gene transcription and translation are not required for the regulation of the process. Rather, post-translational events are involved since the Matrigel-dependent process could be inhibited by the addition of nocodazole, suramin or H7, a protein kinase inhibitor. In contrast, the fibrin matrix-dependent differentiation pathway involved transcriptional and translational events since the addition of either actinomycin D or cycloheximide inhibited this pathway. A modified differential display of RNA extracted from HUVEC after 0, 2, 5, and 24 hours on fibrin revealed expression of a novel cDNA.
Zimrin
A B
AB
Department of Molecular Biology, Holland Laboratory, American Red Cross, Rockville, MD 20855, USA.
Villeponteau
B
B
Maciag
T
T
D001483:Base Sequence; D002454:Cell Differentiation; D002478:Cells, Cultured; D003094:Collagen; D003470:Culture Media; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D004338:Drug Combinations; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects; D005337:Fibrin; D016220:Fibroblast Growth Factor 1 / Q000494:pharmacology; D006801:Humans; D007797:Laminin; D008954:Models, Biological; D008969:Molecular Sequence Data; D009389:Neovascularization, Pathologic; D011509:Proteoglycans; D014158:Transcription, Genetic; D014471:Umbilical Veins
D003470:Culture Media; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D016220:Fibroblast Growth Factor 1; C059485:matrigel; D003609:Dactinomycin; D005337:Fibrin; D003094:Collagen; D003513:Cycloheximide