Unexpected multipotency of melanoblasts isolated from murine skin.
Melanoblasts, precursor of melanocytes, are generated from the neural crest and differentiate into melanocytes during their migration throughout the entire body. The melanoblasts are thought to be progenitor cells that differentiate only into melanocyte. Here, we show that melanoblasts, even after they have already migrated throughout the skin, are multipotent, being able to generate neurons, glial cells, and smooth muscle cells in addition to melanocytes. We isolated Kit-positive and CD45-negative (Kit+/CD45-) cells from both embryonic and neonate skin by flow cytometry and cultured them on stromal cells. The Kit+/CD45- cells formed colonies containing neurons, glial cells, and smooth muscle cells, together with melanocytes. The Kit+/CD45- cells expressed Mitf-M, Sox10, and Trp-2, which are genes known to be expressed in melanoblasts. Even a single Kit+/CD45- cell formed colonies that contained neurons, glial cells, and melanocytes, confirming their multipotential cell fate. The colonies formed from Kit+/CD45- cells retained Kit+/CD45- cells even after 21 days in culture and these retained cells also differentiated into neurons, glial cells, and melanocytes, confirming their self-renewal capability. When the Kit signal was inhibited by the antagonist ACK2, the Kit+/CD45- cells did not form colonies that contained multidifferentiated cells. These results indicate that melanoblasts isolated from skin have multipotency and self-renewal capabilities.
Department of Tissue and Organ Development, Regeneration and Advanced Medical Science, Gifu University Graduate School of Medicine, Gifu, Japan. tmotohas@gifu-u.ac.jp
Tsutomu
T
Motohashi
Katsumasa
K
Yamanaka
Kairi
K
Chiba
Hitomi
H
Aoki
Takahiro
T
Kunisada
D000818:Animals; D000831:Animals, Newborn; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D005434:Flow Cytometry; D007150:Immunohistochemistry; D017493:Leukocyte Common Antigens / Q000378:metabolism; D008544:Melanocytes / Q000166:cytology*; D051379:Mice; D008810:Mice, Inbred C57BL; D039902:Multipotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000166:cytology; D009432:Neural Crest / Q000166:cytology; D009457:Neuroglia / Q000166:cytology; D009474:Neurons / Q000166:cytology; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012867:Skin / Q000166:cytology*
D019009:Proto-Oncogene Proteins c-kit; D017493:Leukocyte Common Antigens