Antigenic immunostaining patterns in somatic hybrids of human HeLa cells and mouse fibroblasts 3T3.4E propagated in conventional medium and delipidized serum.
Somatic cell hybrids were obtained with electric pulse by fusion of human epithelial HeLa cells derived from a carcinoma of the uterine cervix and mouse fibroblasts 3T3.4E, deficient in thymidine kinase. Hybrids were selected and propagated in HAT media; some experiments were carried out in medium with delipidized serum. The hybrid cells were characterized by indirect immunofluorescence with a biotin-streptavidin system using a panel of nine monoclonal antibodies specific for membrane and cytoplasmic antigens of parental cells: intermediate filaments (keratins and vimentin), HLA class 1 (beta 2-microglobulin), cell activation (EGF and transferrin receptors) and cellular adhesion (fibronectin and laminin). All of these antigens were expressed in HeLa cells cultured in conventional medium or with delipidized serum. Conversely mouse fibroblasts contained only vimentin, fibronectin and laminin. All the parental antigens were present in first passage hybrid cells cultured in conventional medium. Vimentin, fibronectin and laminin were maintained in fourth passage hybrids whereas keratins, beta 2-microglobulin, EGF and transferrin receptors were no longer detected. When propagated in medium with delipidized serum, hybrid cells re-expressed these antigens after 5 days of culture. These findings suggest that the reexpression of HeLa cell antigens in hybrid cells was related to deficiency in vitamin A.
Gallagher
T
T
INSERM U 209, CNRS UA 601, Clinique Dermatologique, Hôpital E. Herriot, Lyon, France.
Chardonnet
Y
Y
Viac
J
J
Patet
J
J
Lefèvre
A
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000941:Antigens / Q000276:immunology* / Q000378:metabolism; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology; D066246:ErbB Receptors / Q000276:immunology / Q000378:metabolism; D005347:Fibroblasts / Q000276:immunology* / Q000378:metabolism; D005353:Fibronectins / Q000276:immunology / Q000378:metabolism; D006367:HeLa Cells / Q000166:cytology; D006801:Humans; D006822:Hybrid Cells / Q000276:immunology* / Q000378:metabolism; D007150:Immunohistochemistry; D007621:Karyotyping; D007633:Keratins / Q000276:immunology / Q000378:metabolism; D007797:Laminin / Q000276:immunology / Q000378:metabolism; D008055:Lipids; D008562:Membrane Glycoproteins / Q000276:immunology / Q000378:metabolism; D051379:Mice; D018396:Mucin-1; D011990:Receptors, Transferrin / Q000276:immunology / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000032:analysis / Q000494:pharmacology; D014746:Vimentin / Q000276:immunology / Q000378:metabolism; D001613:beta 2-Microglobulin / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000941:Antigens; D003470:Culture Media; D005353:Fibronectins; D007797:Laminin; D008055:Lipids; D008562:Membrane Glycoproteins; D018396:Mucin-1; D011990:Receptors, Transferrin; D014746:Vimentin; D001613:beta 2-Microglobulin; D012710:Serum Albumin, Bovine; D007633:Keratins; D066246:ErbB Receptors