Technique for the Growth of Epidermal Sheets Obtained from Patients Undergoing Reduction Mammoplasty.

摘要

This report discusses cases in which discarded skin was obtained from patients undergoing reduction mammoplasty. The epidermis was enzymatically separated from the dermis. Keratinocytes were isolated from the epidermis and seeded in 25 cm2 cell culture flasks. The growth medium used was Keratinocyte Growth Medium which has been developed for the growth of keratinocytes. In approximately 2 weeks the primary keratinocyte cultures were nearly confluent and were serially subcultured to expand the volume of cells. When secondary cultures reached confluence, the cell medium was changed Dulbecco's Modified Eagles's medium containing 10% fetal calf serum. The change to a medium containing serum and a higher calcium concentration induced the keratinocytes to stratify into multi-layered sheets. These epidermal sheets were removed from the culture flask with Dispase, a neutral protease and attached to petrolatum gauze. At this point the sheets could be used as skin grafts.