Modulation of normal human melanocyte dendricity by growth-promoting agents.
Dendrite formation and extension, which comprise a characteristic morphology of human normal melanocytes in the skin, represent one of the functional activities of melanocytes, the ability to transfer melanosomes into neighboring keratinocytes. However, the morphology of the melanocyte in vitro is usually quite different from that observed in vivo. it is probably due to the hyperproliferative condition of the melanocytes in culture. No studies have ever compared the effects of a single factor on both dendricity and proliferation at the same time. Therefore, we have compared the effects of six growth-promoting agents commonly used for melanocyte cultures on dendrite formation and proliferation. The addition of agents that increase the intracellular levels of cyclic adenosine monophosphate (cAMP)--dibutyryl cyclic adenosine monophosphate (db cAMP; 1 mM) or isobutylmethyl xanthine (IBMX; 0.1 mM)--had a strong effect on dendrite formation and a negative effect on proliferation. This was especially true with db cAMP. In the presence of 2% or 5% of heat-inactivated fetal bovine serum (FBS), dendrite formation was significantly increased as was proliferation. The number of dendrites was decreased in the culture with 12-o-tetradecanoylphorbol-13-acetate (TPA), but cell growth was slightly increased. With human recombinant basic fibroblast growth factor (bFGF) (0.5, 1.0 ng/ml) in the presence of bovine pituitary extract (BPE) (60 micrograms/ml), cell growth was increased. With 2 ng/ml of bFGF, however, a strong inhibitory effect on proliferation was observed. However, dendrite formation was constant at all concentrations of bFGF tested (0.5, 1.0 or 2.0 ng/ml) with BPE (30 or 60 micrograms/ml). In this study, we have demonstrated that dendrite formation was suppressed by the reagents that stimulate melanocyte proliferation, and vice versa, with the only exception being heat-inactivated FBS. Both dendrite formation and proliferation were induced by the heat-inactivated FBS. This approach is crucial to the development of an adequate culture system for proliferation and/or dendrite formation of normal human melanocytes. It is necessary to keep these aspects in mind as we further investigate the biology of melanocytes, especially the cell-to-cell interactions between melanocytes and keratinocytes, involved in melanogenesis and melanin pigmentation in vivo. This study also provides practical and important information for a future reconstitutive skin system composed of melanocytes, keratinocytes, and fibroblasts in a single culture medium.
Nakazawa
K
K
Skin Substitutes Laboratory, CNRS-URA 1341, Edouard Herriot Hospital, Lyon, France.
Damour
O
O
Collombel
C
C
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D002417:Cattle; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000502:physiology; D003712:Dendrites / Q000648:ultrastructure*; D005312:Fetal Blood; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006133:Growth Substances / Q000494:pharmacology*; D006358:Hot Temperature; D006801:Humans; D008544:Melanocytes / Q000648:ultrastructure*; D011994:Recombinant Proteins / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D006133:Growth Substances; D011994:Recombinant Proteins; D016222:Fibroblast Growth Factor 2; D003994:Bucladesine; D000242:Cyclic AMP; D013755:Tetradecanoylphorbol Acetate; D015056:1-Methyl-3-isobutylxanthine