A new spectrophotometric method for simple quantification of melanosomal transfer from melanocytes to keratinocytes

作者：Kasraee Behrooz^*; Pataky Marcela; Nikolic Damjan S; Carraux Pierre; Piguet Vincent; Salomon Denis; Sorg Olivier; Saurat Jean Hilaire

来源：Experimental Dermatology, 2011, 20(11): 938-942.

DOI：10.1111/j.1600-0625.2011.01356.x

• cell culture
• keratinocytes
• melanocytes
• melanosome transfer
• pigmentation

Three major difficulties must be overcome to establish a quantitative method for melanosomal transfer analysis: (i) establishing a three-dimensional co-culture reassuring direct melanocyte to keratinocyte transfer, (ii) separation of melanocytes and keratinocytes following co-culture and (iii) melanosome quantification in each cell population. Melanocytes and keratinocytes are cultured on the opposite sides of the porous membrane of hanging cell inserts (1 mu m pores, 2 x 10(6) pores/cm(2)). Cell separation is performed after 3 days of co-culture by simple trypsinisation. Melanosome quantification in separated cell populations was accomplished by an ELISA-like method using gp-100 as the antigen. Melanocytes and keratinocytes come into 'direct' contact through the pores, and melanosomal transfer is accomplished without cell passage through the membrane. Cell separation by simple trypsinisation results in pure melanocyte and keratinocyte populations. Melanosome quantification by the ELISA-like method proved to be sensitive and specific to distinguish the known inhibitors and inducers of melanosomal transfer.

• 出版日期2011-11