Stem Cell Factor and/or Endothelin‐3 Dependent Immortal Melanoblast and Melanocyte Populations Derived From Mouse Neural Crest Cells

Kawa, Yoko; Ito, Masaru; Ono, Hirotake; Asano, Mari; Takano, N.; Ooka, Seido; Watabe, Hidenori; Hosaka, Eri; Baba, Takao; Kubota, Yasuo; Mizoguchi, Masahiro

doi:10.1034/j.1600-0749.13.s8.14.x

Cited by 40 publications

(60 citation statements)

References 13 publications

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“…The cells were then reacted with alkaline phosphatase-labeled anti-rabbit IgG (at 100 Â dilution; DAKO) for 1 h at RT. Colorization was developed using the new fuchsin substrate system (DAKO) and levamisole solution (Vector, Burlingame, CA) as an inhibitor of internal alkaline phosphatase (Kawa et al, 2000). Positive cells which possessed reaction products were estimated as ( positive).…”

Section: Immunocytochemical Stainingmentioning

confidence: 99%

Stimulation of the proliferation and differentiation of mouse pink‐eyed dilution epidermal melanocytes by excess tyrosine in serum‐free primary culture

Hirobe

Wakamatsu

Ito

et al. 2002

Journal Cellular Physiology

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The epidermal cell suspensions of the neonatal dorsal skin derived from wild type mouse at the pink-eyed dilution (p) locus (black, C57BL/10JHir-P/P) and their congenic mutant mouse (pink-eyed dilution, C57BL/10JHir-p/p) were cultured with a serum-free melanocyte growth medium supplemented with additional L-tyrosine (Tyr) from initiation of the primary culture. L-Tyr inhibited the proliferation of P/Pmelanocytes in a dose-dependent manner, whereas L-Tyr stimulated the proliferation of p/p melanoblasts and melanocytes regardless of dose. On the other hand, L-Tyr stimulated (P/P) or induced (p/p) the differentiation of epidermal melanocytes in a dose-dependent manner. In both P/P and p/p melanoblasts and melanocytes cultured with 2.0 mM L-Tyr for 14 days, slight increases in contents of eumelanin marker, pyrrole-2,3,5-tricarboxylic acid (PTCA) and pheomelanin marker, aminohydroxyphenylalanine (AHP) were observed. The average number of total melanosomes (stages I, II, III, and IV) per P/P melanocyte was not changed by L-Tyr treatment, but the proportion of stage IV melanosomes in the total melanosomes was increased. On the contrary, in p/p melanoblasts and melanocytes L-Tyr increased dramatically the number of stage II, III, and IV melanosomes as well as the proportion of stage III melanosomes. Contents of PTCA and eumelanin precursor, 5,6-dihydroxyindole-2-carboxylic acid (DHICA) of cultured media in p/p melanocytes were much more greatly increased than in P/P melanocytes. However, contents of AHP and pheomelanin precursor, 5-S-cysteinyldopa (5-S-CD) of cultured media in p/p melanocytes were increased in a similar tendency to P/Pmelanocytes. These results suggest that p/p melanocytes in the primary culture are induced to synthesize eumelanin by excess L-Tyr, but difficult to accumulate them in melanosomes.

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Section: Immunocytochemical Stainingmentioning

confidence: 99%

Stimulation of the proliferation and differentiation of mouse pink‐eyed dilution epidermal melanocytes by excess tyrosine in serum‐free primary culture

Hirobe

Wakamatsu

Ito

et al. 2002

Journal Cellular Physiology

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show abstract

“…The cells were then reacted with alkaline phosphatase-labeled anti-rabbit IgG (at 100× dilution; DAKO) for 1 hr at RT. Colorization was developed using the new fuchsin substrate system (DAKO) and levamisole solution (Vector, Burlingame, CA) as an inhibitor of internal alkaline phosphatase (Kawa et al, 2000).…”

Section: Immunocytochemical Stainingmentioning

confidence: 99%

Effects of genic substitution at the pink‐eyed dilution locus on the proliferation and differentiation of mouse epidermal melanocytes in vivo and in vitro

et al. 2002

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Cells positive to the dopa reaction (melanocytes) as well as to the combined dopa-premelanin reaction (melanoblasts and melanocytes) in the epidermis of C57BL/10JHir-p/p (pink-eyed dilution) mice were fewer and less reactive than in C57BL/10JHir (black, P/P) mice, suggesting that the proliferation and differentiation of p/p melanocytes are inhibited. To confirm the inhibitory effects of p gene on the proliferation and differentiation of epidermal melanocytes, we cultured epidermal cell suspensions of neonatal skins from P/P and p/p in a serum-free medium. The proliferation and differentiation of p/p melanoblasts/melanocytes in primary culture were greatly inhibited as compared to P/P melanoblasts/melanocytes. The morphology of p/p melanoblasts/melanocytes cultured in melanocyte growth medium, though non-pigmented, was similar to P/P melanocytes; namely, dendritic, polygonal, or epithelioid. About 8% of p/p cells cultured in melanocyte growth medium were positive to the dopa reaction, and about 25% were reactive to the combined dopa-premelanin reaction. Eumelanin content in p/p was extremely reduced compared to P/P. The immunocytochemical staining of p/p melanoblasts/melanocytes revealed that they are negative to tyrosinase, but reactive to tyrosinase-related protein (TRP)-1, TRP-2, and c-kit. However, the reactivities in p/p were lower than in P/P. Although the differentiation of p/p melanoblasts was not induced by endothelin (ET)-1, ET-2, and ET-3, the proliferation of p/p melanoblasts was stimulated by them. These results suggest for the first time that p gene exerts its influence on the proliferative activities of mouse epidermal melanoblasts by affecting the regulatory mechanisms dependent on the function of ETs.

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“…NCCmelb4 cells were cultured in Eagle's modified essential medium (Eagle's MEM, IBL, Maebashi, Japan) supplemented with 5% fetal bovine serum (FBS, GIBCO, Grand Island, NY, USA) and 50 ng/ml of recombinant mouse stem cell factor (SCF, kindly supplied by Kirin Brewery, Tokyo, Japan). NCC-E3 cells were induced from primary NCC cultures as described previously [9]. NCC-E3 cells were cultured in Eagle's MEM supplemented with 15% FBS and 100 nM endothelin 3 (EDN3, Peptide Institute Inc, Osaka, Japan).…”

Section: Cells and Culture Conditionsmentioning

confidence: 99%

“…After three further washes with TBS-T, the immunoreactivities of antibodies were detected using an ECL + Western Blotting Detection System (Amersham Biosciences) according to the manufacturer's instructions. Electron microscopy and DOPA reactions were performed as described previously [9].…”

Section: Western Blotting Electron Microscopy and Dopa Reactionmentioning

confidence: 99%

“…The first of those NCC-derived cells, named NCC-E3 cells, have melanosomes most of which display well organized structures and exhibit lumenal fibrillar striations, indicating that they are Stage II melanosomes. NCC-E3 cells are positive for the L-3,4-dihydroxyphenylalanine (DOPA) reaction, and for Tyrp1, Dct, tyrosinase, Kit and endothelin B receptor (Ednrb) [9]. The second type of cells, named NCCmelb4, are immature melanocyte precursors.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Proteomic analysis of immature murine melanocytes at different stages of maturation: A crucial role for calreticulin

Kawase

Kushimoto

Kawa

et al. 2008

Journal of Dermatological Science

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Stem Cell Factor and/or Endothelin‐3 Dependent Immortal Melanoblast and Melanocyte Populations Derived From Mouse Neural Crest Cells

Cited by 40 publications

References 13 publications

Stimulation of the proliferation and differentiation of mouse pink‐eyed dilution epidermal melanocytes by excess tyrosine in serum‐free primary culture

Stimulation of the proliferation and differentiation of mouse pink‐eyed dilution epidermal melanocytes by excess tyrosine in serum‐free primary culture

Effects of genic substitution at the pink‐eyed dilution locus on the proliferation and differentiation of mouse epidermal melanocytes in vivo and in vitro

Proteomic analysis of immature murine melanocytes at different stages of maturation: A crucial role for calreticulin

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