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Endothelial cells are indispensable components of the vascular system, and play pivotal roles during development and in health and disease. Their properties have been studied extensively byin vivo analysis of genetically modified mice. | ễFEBS Journal Development of a new method for isolation and long-term culture of organ-specific blood vascular and lymphatic endothelial cells of the mouse Takashi Yamaguchi Taeko Ichise Osamu Iwata Akiko Hori Tomomi Adachi Masaru Nakamura Nobuaki Yoshida and Hirotake Ichise Laboratory of Gene Expression and Regulation Center for ExperimentalMedicine Institute of MedicalScience University of Tokyo Japan Keywords Cre loxP recombination endothelial cell culture endothelialheterogeneity SV40 tsA58 large T antigen transgenic mouse Correspondence H. Ichise Laboratory of Gene Expression and Regulation Center for Experimental Medicine Institute of MedicalScience University of Tokyo 4-6-1 Shirokanedai Minato-ku Tokyo 108-8639 Japan Fax 81 3 5449 5455 Tel 81 3 5449 5754 E-mail h-ichise@ims.u-tokyo.ac.jp Received 27 November 2007 revised 13 February 2008 accepted 22 February 2008 doi 10.1111 j.1742-4658.2008.06353.x Endothelial cells are indispensable components of the vascular system and play pivotal roles during development and in health and disease. Their properties have been studied extensively by in vivo analysis of genetically modified mice. However further analysis of the molecular and cellular phenotypes of endothelial cells and their heterogeneity at various developmental stages in vascular beds and in various organs has often been hampered by difficulties in culturing mouse endothelial cells. In order to overcome these difficulties we developed a new transgenic mouse line expressing the SV40 tsA58 large T antigen tsA58T Ag under the control of a binary expression system based on Cre loxP recombination. tsA58T Ag-positive endothelial cells in primary cultures of a variety of organs proliferate continuously at 33 C without undergoing cell senescence. The resulting cell population consists of blood vascular and lymphatic endothelial cells which could be separated by immunosorting. Even when cultured for two months the cells maintained endothelial cell properties as .
