Evidence suggests that human cytomegalovirus is resident in the kidneys of seropositive donors at the time of transplantation, and CMV has been implicated in both glomerulonephritis and interstitial nephritis. In this study we assessed the interactions of CMV with two human renal cell types in culture: glomerular visceral epithelial cells (GVE) and renal tubular epithelial (RTE) cells. GVE permitted viral adsorption, penetration, nuclear translocation, and restricted viral transcription. However, early viral protein expression was not detectable by immunofluorescence and infectious virions were not produced. In contrast, retinoic acid-treated GVE permitted early viral protein expression and supported CMV replication. RTE also permitted viral adsorption and penetration. CMV-specific early proteins were readily observed by immunofluorescence, and CMV DNA replication was observed by DNA dot blot hybridization. Assays comparing viral yield with viral DNA synthesis indicated that RTE were capable of supporting persistent and prolonged viral expression without significant cell death for at least 55 days after infection. We believe that these findings should explain chronic viruria in individuals with symptomatic and asymptomatic CMV infection. In addition, GVE could also be a potential source of CMV transmission when altered by disease or transplantation.