IMMORTALIZATION OF MULTIPLE CELL-TYPES FROM TRANSGENIC MICE USING A TRANSGENE CONTAINING THE VIMENTIN PROMOTER AND A CONDITIONAL ONCOGENE

Differentiated clonal cell lines were obtained from transgenic mice ca rrying a recombinant gene composed of DNA coding for a temperature-sen sitive mutant of the simian virus large T antigen under the control of regulatory elements of the human vimentin gene. In response to mitoge nic factors the vimentin promoter is activated in the presence of seru m in almost all cultured cells independently of their origin. The expr ession of the T antigen could be controlled both at the level of trans cription since the vimentin promoter is growth-regulated and at the le vel of the protein structure through the temperature stability of the T antigen. Indeed, the switch-off of the oncogene protein is obtained by serum deprivation of the culture and achieved with enhancement of t he growth temperature. From transgenic mice several types of clonal di fferentiated cell lines were established and characterized including m elanocytes, macrophages, mesangial, muscle, and endothelial cells. Mel anocytes displayed melanin while endothelial cells from brain and hear t expressed the related factor VIII and low density lipoprotein absorp tion capacities. Mesangial cells from kidney exhibited numerous desmos omes. Typical markers of macrophages from bone marrow were observed wh ile skeletal muscle cells fused and contracted. (C) 1994 Academic Pres s, Inc.