Abstract
Human primary thyroid follicular epithelial cells were transfected with a plasmid containing an origin-defective SV40 genome (SVori-) to produce several immortal cell lines. Two of the 10 cell lines analysed expressed specific features of thyroid epithelial function (iodide-trapping and thyroglobulin production). These two lines were characterised in detail and found to be growth factor-independent, capable of anchorage-independent growth at low frequency but non-tumorigenic in nude mice. These differentiated, These differentiated, partially transformed cell lines were shown to be suitable for gene transfer at high frequency using simple coprecipitation techniques.
Highlights
MethodsThese were prepared as we have previously described (Williams et al, 1987)
The majority of human malignancies are epithelial in origin (Parkin & Muir, 1988), but understanding of the steps leading to the neoplastic transformation of human epithelial cells has been hampered by several problems
Human cells are recognised to be more resistant than rodent cells to in vitro transformation, whether spontaneous or induced by various means (DiPaolo, 1983; Sager, 1984) and epithelial cells of human origin appear to be even more resistant than mesenchymal derivatives such as fibroblasts, as illustrated by studies of SV40-induced transformation (Girardi et al, 1965; Chang, 1986)
Summary
These were prepared as we have previously described (Williams et al, 1987). Dissected from lobectomy specimens (surgically removed for solitary thyroid nodules) and digested with a mixture of collagenase (100 IU ml- ') and dispase (1 mg ml- ') and the follicles were pooled, filtered through a 200 1Lm nylon mesh and washed with Hank's calcium- and magnesium-free balanced salt solution (HBSS). The follicular epithelium was plated as monolayers of 5 x 105 cells per 60 mm dish in RPMI 1640 medium with 10% fetal calf serum and 0.1 mU ml' bTSH (bovine thyrotropic hormone; Sigma). Thyroid follicular epithelium from one female patient aged 35 years was used for these cultures
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