A new method for culturing human hair follicle cells without a biological feeder layer has been developed. Dispersed hair follicle cells obtained from plucked hairs have been successfully grown on floating mixed collagen membranes (collagen types I and IV) and have reached a confluent state. They could be subcultured up to twice. Differentiation of the cells was maintained throughout a month in culture. Stratification of the cells and continuous epithelial pavement formation were seen, as in cultured epidermal keratinocytes.