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Keratinocytes in culture accumulate phagocytosed melanosomes in the perinuclear area
Author(s) -
Ando Hideya,
Niki Yoko,
Yoshida Masaki,
Ito Masaaki,
Akiyama Kaoru,
Kim JinHwa,
Yoon TaeJin,
Lee JeungHoon,
Matsui Mary S.,
Ichihashi Masamitsu
Publication year - 2010
Publication title -
pigment cell and melanoma research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.618
H-Index - 105
eISSN - 1755-148X
pISSN - 1755-1471
DOI - 10.1111/j.1755-148x.2009.00640.x
Subject(s) - melanosome , keratinocyte , phagocytosis , microbiology and biotechnology , in vitro , biology , chemistry , melanin , biochemistry
Summary There are many techniques for evaluating melanosome transfer to keratinocytes but the spectrophotometric quantification of melanosomes incorporated by keratinocyte phagocytosis has not been previously reported. Here we describe a new method that allows the spectrophotometric visualization of melanosome uptake by normal human keratinocytes in culture. Fontana‐Masson staining of keratinocytes incubated with isolated melanosomes showed the accumulation of incorporated melanosomes in the perinuclear areas of keratinocytes within 48 h. Electron microscopic observations of melanosomes ingested by keratinocytes revealed that many phagosomes containing clusters of melanosomes or their fragments were localized in the perinuclear area. A known inhibitor of keratinocyte phagocytosis which inhibits protease‐activated receptor‐2, i.e., soybean trypsin inhibitor, decreased melanosome uptake by keratinocytes in a dose‐dependent manner. These data suggest that our method is a useful model to quantitate keratinocyte phagocytosis of melanosomes visually in vitro.