Behavioral differences between donor site-matched adult and neonatal melanocytes in culture

Abstract

Little is known about the biologic behaviors of cultured melanocytes in relation to donor age. To investigate age-dependent differences, neonatal and adult melanocytes were isolated from the same anatomical site, the foreskin, and cultured in the same growth medium supplemented with cAMP inducers (choleratoxin and 3-isobutyl-methylxanthine). The morphology, melanin content, pattern of melanosome distribution, and growth rate were then compared. Neonatal melanocytes were bipolar in appearance, whereas adult melanocytes were highly dendritic in appearance. Image analysis showed that adult melanocytes were larger and longer, and had a greater number of dendrites than neonatal melanocytes. When the growth medium was replaced by a medium without cAMP inducers, adult melanocytes showed a change in their morphology from dendritic to spindle-shaped, while the morphology of neonatal melanocytes remained unchanged. Melanosomes of adult melanocytes were distributed singly along the dendrites, and extracellular secretion of melanosomes was also found. In contrast, melanosomes of neonatal melanocytes were aggregated near the nuclei. No age-dependent differences in melanin content and growth rate were noted in the donor sitematched cultured melanocytes. These results suggest that donor age is one of the factors involved in determining melanocyte dendricity and melanosome distribution, and that increased dendricity of adult melanocytes is due to increased sensitivity to cAMP inducers. In addition, the adult melanocytes established in our culture system, which resembled dendritic melanocytes in vivo, could be considered a desirable model for studying the mechanisms of adult-onset hyperpigmentary disorders and melanogenesis. Key words Dendricity · Melanosome distribution · cAMP · Donor ageAbbreviations bFGF basic fibroblast growth factor · cAMP adenosine 3′,5′-cyclic monophosphate · CT choleratoxin · IBMX 3-isobutyl-methylxanthine · PBS phosphate-buffered saline · TPA 12-O-tetradecanoylphorbol-13-acetate

Chronic Stress Suppresses the Expression of Cutaneous Hypothalamic–Pituitary–Adrenocortical Axis Elements and Melanogenesis

“…Normal human epidermal melanocytes were derived from young male adult foreskins (ethnic Han/aged 18–22 years) obtained at circumcision following standard protocols [41]. Melanocytes were incubated with MCDB153 medium (Sigma-Aldrich, USA) containing 1 nM choleratoxin (Sigma-Aldrich, USA), 0.1 mM 3-isobutyl-1-methylxanthine (Sigma-Aldrich, USA), 1.6 nM phorbolesters (Sigma-Aldrich, USA), 5 µg/mL insulin (Sigma-Aldrich, USA) and incubated at 37°C in a humidified atmosphere containing 5% CO 2 .…”

Interleukin 10 protects primary melanocyte by activation of Stat-3 and PI3K/Akt/NF-κB signaling pathways

“…Normal human foreskin-derived epidermal melanocytes (NHEM) were derived from young male adult foreskins (ethnic Han/aged 18-22 years) obtained at circumcision following standard protocols [29]. Briefly, foreskins were cut into strips and digested with 0.25% trypsin at 4°C for 20 h. Epidermis was separated from dermis.…”

Donor Specific Response of Estrogen and Progesterone on Cultured Human Melanocytes

“…Normal human melanocytes were derived from the adult foreskin as previously described (7) and maintained in a growth medium consisting of MCDB-153, 4% heat-inactivated fetal bovine serum (FBS) (Gibco BRL, Grand Island, NY, U.S.A.), 10-7 M -melanocyte stimulating hormone ( -MSH) (Sigma Chemical Co., St. Louis, MO, U.S.A.), 8 nM 12-O-tetradecanoyl phorbol 13-acetate (TPA) (Sigma Chemical Co., St. Louis, MO, U.S.A.), 0.6 ng/mL human recombinant basic fibroblast growth factor (FGF) (Calbiochem, San Diego, CA, U.S.A.), 5 g/mL insulin (Sigma Chemical Co., St. Louis, MO, U.S.A.), 1 g/mL -tocopherol (Sigma Chemical Co., St. Louis, MO, U.S.A.), and 1% penicillin-streptomycin (10,000 units/mL and 10,000 g/ mL, respectively) (Sigma Chemical Co., St. Louis, MO, U.S.A.).…”