Conditional immortalization of growth factor-responsive cardiac endothelial cells from H-2K(b)-tsA58 mice.
Although mouse endothelial cells (EC) may advance our understanding of endothelial function, primary EC remain difficult to isolate. We have established a murine cardiac endothelial cell line (MCEC-1) from transgenic mice harboring a temperature-sensitive simian virus 40 large TAg gene (tsA58 TAg) under H-2K(b) class I promoter control. MCEC-1 cells were characterized by their ability to form tubes, Griffonia simplicifolia isolectin B4 binding, and CD31, intercellular adhesion molecule (ICAM)-2, and endoglin expression. MCEC-1 cells proliferated rapidly under permissive conditions [33 degrees C with interferon (IFN)-gamma], where the T antigen is active and transcription is activated by the presence of IFN-gamma, whereas under nonpermissive conditions (38 degrees C without IFN-gamma) proliferation was reduced by 30-fold and the EC showed enhanced proliferation in response to growth factors. Expression of E- and P-selectin, ICAM-1, and vascular cell adhesion molecule-1 was upregulated by tumor necrosis factor-alpha and interleukin-1 beta, and MCEC-1 cells, in contrast to primary EC, were amenable to transfection by lipofection. This novel line will allow further study of the role of the endothelium in cardiovascular disease. Moreover, this technique will allow EC to be readily obtained from genetically modified mice backcrossed with H-2K(b)-tsA58 mice.
Lidington
Elaine A
EA
British Heart Foundation Cardiovascular Medicine Unit, National Heart and Lung Institute, London, United Kingdom.
Rao
Ravi M
RM
Marelli-Berg
Federica M
FM
Jat
Parmjit S
PS
Haskard
Dorian O
DO
Mason
Justin C
JC
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics*; D000970:Antineoplastic Agents / Q000494:pharmacology; D015815:Cell Adhesion Molecules / Q000235:genetics; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D003326:Coronary Circulation / Q000502:physiology*; D003327:Coronary Disease / Q000503:physiopathology; D004730:Endothelium, Vascular / Q000166:cytology* / Q000502:physiology*; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D006133:Growth Substances / Q000494:pharmacology*; D007375:Interleukin-1 / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D013696:Temperature; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D000952:Antigens, Polyomavirus Transforming; D000970:Antineoplastic Agents; D015815:Cell Adhesion Molecules; D006133:Growth Substances; D007375:Interleukin-1; D014409:Tumor Necrosis Factor-alpha