Immortalization of precursors of endodermal, neuroectodermal and mesodermal lineages, following the introduction of the simian virus (SV40) early region into F9 cells

Abstract F9 embryonal carcinoma cells were transfected with a hybrid plasmid containing the early genes of the simian virus SV40 under the control of the adenovirus type 5 E1 A promoter [21]. These cells were induced to differentiate in aggregates in the presence of retinoic acid (RA). Unlike the derivatives of F9 that are usually obtained in this manner, the plasmid-containing cells were both programmed and immortalized; in addition, expression of the SV40 T antigen was now triggered. These immortalized cells could be separated into three classes: (1) extraembryonic derivatives, (2) embryonic differentiated tissues, (3) immature cells surrounding the differentiated cells. When injected into mice, the mixture of these cells gave rise to multipotential tumors. From the immature cells, committed precursors of the neuroectodermal, endodermal, and mesodermal pathways could be isolated by cloning and selection according to: (a) their specific pattern of differentiation in the tumors and (b) the occurrence of specific markers in the differentiated progeny. The isolation of stable immortalized cell lines corresponding to precursors of the three primitive germ layers and capable of differentiating reproducibly along a particular restricted pathway should facilitate molecular studies on early embryonic development in mouse.