Establishment of a cell line retaining Langerhans cell characteristics

A discontinuous gradient of polyethylene glycol (PEG) with dimethyl sulfoxide (DMSO) was used to induced fusion between guinea pig epidermal cells (GPEC) enriched in Langerhans cells (LC) and cells of a human fibrosarcoma line (T-1080) deficient in hypoxanthine phosphoribosyl transferase (HPRT). The rapidly proliferating carrier T-1080 cells lack plasma membrane ATPase activity, Fc receptors, C3b receptors, guinea pig Ia-antigens, and nonspecific esterase activity, all characteristic of LC, and absent on other guinea pig epidermal cells. GPE/T-1080 hybrid cells were selected in hypoxanthine/aminopterin/thymidine (HAT) medium and by C3b and Fc rosette formation. A resultant continuous cell line was characterized karyotypically, histochemically, ultrastructurally and immunologically.