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Cellosaurus publication CLPUB00265

Publication number CLPUB00265
Authors Stoner G.D., Kaighn M.E., Resau J.H., Naito Z., Bowman D., Harris C.C.
Title Immortalized human esophageal epithelial cell lines for studies of carcinogen-induced cell transformation.
Citation In Vitro Cell. Dev. Biol. 26:23A-23A(1990)
Web pages https://www.jstor.org/stable/4296437
Abstract Treatment of normal human epithelial cells with chemical carcinogens has rarely led to neoplastic cell transformation. One approach to the development of human epithelial cell cultures for studies of carcinogen- induced cell transformation is to immortalize the cells with certain viral genes, such as the SV40 early region genes. In the present study, normal human esophagus tissue obtained from autopsy was explanted in serum-free LHC-9 medium. The epithelial outgrowths were subcultured, then transfected by strontium phosphate coprecipitation with plasmid pRSV-T consisting of the RSV-LTR promoter and the sequence encoding the SV40 large T-antigen The transfected cells but not the sham-transfected controls formed multilayered colonies within 2-3 weeks which grew exponentially for 8-10 weeks, then senesced After a "crisis" of 6-8 months, growth resumed in isolated colonies. Two lines (HET-1A and HET-2A), were developed and have now undergone 137 and 193 population doublings, respectively Both have an epithelial morphology, stain for cytokeratins and the SV40 T antigen gene by immunofluorescence, and have remained nontumorigenic in nude, athymic mice for 12 months. Karyotypic analysis by Giemsa banding has shown that HET-1A is hypodiploid (34-40 chromosomes), whereas HET-2A is hypotriploid (57-64 chromosomes). Both cell lines are sensitive to serum and TGF- induced terminal cell differentiation. Clones of both lines have been adapted to grow in either EGF-, insulin-, or bovine pituitary extract-free medium. This serum and growth factor-free system and the 2 immortalized esophageal cell lines should be useful for studies of carcinogen-induced cell transformation.
Cell lines CVCL_3702; HET-1A