[Methods for optimized cultivation of hair cells from C3H mice]

Methods for the optimized culturing the hair cells from 4-day-old C3H mice are described. The skin was obtained and soaked in 500 unit/ml dispase at 4 degrees C for 24 hr. Then the epidermis was removed and the dermis was further treated with 0.25% collagenase for 1 hr at 37 degrees C. The hair roots isolated by collagenase digestion of the dermis were dispersed by the treatment with the mixture of trypsin and EDTA into a cell suspension and plated on a substrate. Attachment of hair cells was dependent on serum concentrations and enhanced by collagen coating of the surface of dishes. MCDB 153 was beneficial for the growth of hair cells and discouraged for the growth of dermal fibroblasts. The cells could not grow at inoculum size of less than 3 x 10(4) cells/cm2. Addition of a crude bovine pituitary extract in MCDB 153 had the greatest impact on hair cell growth. Functional integrity of the cultured hair cells were maintained, since the hair-specific cytoskeletal proteins were expressed in these cells under the present experimental conditions. These desirable conditions could allow selective growth of the hair cells and provide the model system which could be used for the study of hair growth.