Enhanced cell survival of melanocyte spheroids in serum starvation condition

Autologous melanocyte transplantation for vitiligo treatment by use of melanocyte suspension has drawbacks including cell damage in cell preparation and transportation, difficult manipulation and low engraftment rate in acral vitiligious lesions. We have proposed the concept of cellular patch as an alternative solution. In the development of melanocyte patches, we have shown that chitosan membrane supports the growth and phenotype expression of melanocytes. Surprisingly, melanocytes spontaneously grow into three-dimensional spheroids on chitosan-coated surface. In this work, we demonstrate that, compared with monolayered melanocytes, melanocyte spheroids show a better survival in growth factor and serum-deprived condition. Survival of melanocytes is further ameliorated when a greater portion of melanocytes is precultured into spheroidal morphology. Melanocyte spheroids disintegrate and the cells return to a physiological dendritic morphology after they are reinoculated on collagen I-coated surface. Our results show that melanocytes are morphologically transformable depending on the substratum used and spheroidal melanocytes have a superior survival to that of monolayered dendritic melanocytes in stringent conditions. Preculturing melanocytes into spheroids can provide melanocytes a survival advantage. Chitosan-based melanocyte patch can be a promising method to enhance the engraftment rate and facilitate the cell preparation and transplantation procedures in melanocyte transplantation for vitiligo treatment.