Renal allograft recipients are at greatly increased risk of developing squamous cell carcinomas. As these frequently arise adjacent to areas of multiple viral warts, we have investigated a possible role for human papillomavirus in malignant transformation within this population. We established, as primary cultures, keratinocytes from 24 lesions of varying degrees of squamous atypia from 9 patients. Ten of 14 cultures screened for the presence of episomal human papillomavirus DNA were positive using a mixed probe for cutaneous human papillomaviruses, although episomal copy was universally lost with continued passage. Three cultures, 2 of which were derived from malignant tissue and 1 from a benign lesion, were positive when screened with a probe for potentially oncogenic human papillomavirus DNAs 5 or 8. Both positive cultures of malignant origin exhibited extended lifespan and have been briefly characterized by morphology and growth requirements.