Comparison of gene expression in bovine aortic endothelium in vivo versus in vitro. Differences in growth regulatory molecules.
Many studies of the endothelium have assumed equivalence between cultured confluent cells and an endothelial lining in vivo. We compared gene expression of bovine aortic endothelial cells (BAECs) in culture versus freshly isolated cells from bovine aortas. Our technique of harvesting in vivo tissue yielded cells that were endothelial by the criteria of their containing von Willebrand factor (vWF) and lacking smooth muscle alpha-actin, by both immunocytochemistry and mRNA analyses. We found that several genes are overexpressed when BAECs are placed into culture, including basic fibroblast growth factor, platelet-derived growth factor B-chain, and thrombospondin. On the other hand, message for vWF is highly expressed in vivo and at lower levels in confluent cultures. The transcripts for transforming growth factor-beta, plakoglobin, and flg (fms-like gene, FGF receptor-1) are comparable in vitro and in vivo. These results demonstrate that significant changes in gene expression occur in the transition from in vivo conditions to tissue culture of endothelial cells. Studies of in vitro endothelium may poorly reflect a quiescent endothelial lining in vivo but may be more similar to cells responding to injury or angiogenic stimuli.
Liaw
L
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Department of Pathology, University of Washington, Seattle 98195.
Schwartz
S M
SM
D000199:Actins / Q000032:analysis / Q000235:genetics; D000818:Animals; D000906:Antibodies; D001011:Aorta / Q000166:cytology; D015152:Blotting, Northern; D002417:Cattle; D002478:Cells, Cultured / Q000737:chemistry; D003598:Cytoskeletal Proteins / Q000032:analysis / Q000235:genetics; D004247:DNA / Q000032:analysis; D051180:Desmoplakins; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D016222:Fibroblast Growth Factor 2 / Q000032:analysis / Q000235:genetics / Q000276:immunology; D000091344:Filaggrin Proteins; D015870:Gene Expression / Q000502:physiology*; D006801:Humans; D007150:Immunohistochemistry; D009131:Muscle, Smooth, Vascular / Q000737:chemistry; D010980:Platelet Membrane Glycoproteins / Q000235:genetics; D010982:Platelet-Derived Growth Factor / Q000032:analysis / Q000235:genetics; D012313:RNA / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D019699:Thrombospondins; D014158:Transcription, Genetic; D016212:Transforming Growth Factor beta / Q000032:analysis / Q000235:genetics; D014471:Umbilical Veins / Q000737:chemistry; D015854:Up-Regulation / Q000235:genetics; D051185:gamma Catenin; D014841:von Willebrand Factor / Q000032:analysis / Q000235:genetics
D000199:Actins; D000906:Antibodies; D003598:Cytoskeletal Proteins; D051180:Desmoplakins; C000717249:FLG protein, human; D000091344:Filaggrin Proteins; D010980:Platelet Membrane Glycoproteins; D010982:Platelet-Derived Growth Factor; D012333:RNA, Messenger; D019699:Thrombospondins; D016212:Transforming Growth Factor beta; D051185:gamma Catenin; D014841:von Willebrand Factor; D016222:Fibroblast Growth Factor 2; D012313:RNA; D004247:DNA