Large-scale cultivation of human melanocytes using collagen-coated Sephadex beads (cytodex 3)

Pure melanocytes were obtained from the epidermis of human foreskin by a modification of a previously described method in which geneticin was added for selective killing of fibroblasts. Purity of the culture was confirmed by light and electron microscopy and by the use of a monoclonal antibody NKI-beteb, which is specific for a vesicular membrane antigen present on melanocytes. Melanocytes were tested for their affinity to several microcarriers. They attached to cytodex 1 and 3 and dorma cell, but they did not attach to glass and gelatin beads. The best results were obtained with cytodex 3. After an almost immediate and total attachment of melanocytes a fourfold to fivefold increase in cell number was achieved on this microcarrier within 3 weeks. With the results obtained, it seems that the collagen-coated cytodex 3 microcarrier surface supports the growth of melanocytes. Preliminary results obtained with a microcarrier cell culture fermenter clearly indicate that the large-scale cultivation of normal human melanocytes in such an automated system is possible.