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Am J Physiol Cell Physiol 282: C67-C74, 2002;
0363-6143/02 $5.00
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Vol. 282, Issue 1, C67-C74, January 2002

Conditional immortalization of growth factor-responsive cardiac endothelial cells from H-2Kb-tsA58 mice

Elaine A. Lidington1, Ravi M. Rao1, Federica M. Marelli-Berg2, Parmjit S. Jat3,4, Dorian O. Haskard1, and Justin C. Mason1

1 British Heart Foundation Cardiovascular Medicine Unit, National Heart and Lung Institute and 2 Department of Immunology, Imperial College School of Technology and Medicine, Hammersmith Hospital, London W12 ONN; 3 Ludwig Institute for Cancer Research, University College Medical School, London W1W 7BS; and 4 Department of Biochemistry and Molecular Biology, University College London, London WC1E 6BT, United Kingdom

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-2Kb 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°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°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-1beta , 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-2Kb-tsA58 mice.

endothelium; proliferation; adhesion; Immortomouse


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