Novel method for isolating human melanoblasts from keratinocyte culture.
The characterization of melanoblasts is important for understanding their in vivo development, melanoma formation, and pigment-related disorders. However, no methods have been reported for the isolation of melanoblasts from human skin. Using a 'calcium-pulse' technique, involving the differentiation of human keratinocytes with high calcium and the subsequent spontaneous separation of the epidermal sheets, we effectively isolated human melanoblasts (keratinocyte-adapted melanoblasts, KaMBs) from keratinocyte culture. The KaMBs expressed early melanogenesis-related genes, including BRN2, which is a known melanoblast marker. Moreover, the KaMBs displayed much higher proliferative and growth capacities than the primary melanocytes. Considering that keratinocytes might provide an in vivo-like environment for KaMBs during isolation and in vitro culture, the 'calcium-pulse' technique offers an unprecedented, easy, and efficient method for the isolation of human melanoblasts, retaining the original characteristics of these cells.
Bioscience Research Division, R&D Unit, AmorePacific Corporation, Yongin, Gyeonggi-do, Korea.
Eun-Gyung
EG
Cho
Bum-Ho
BH
Bin
Hyunjung
H
Choi
Phil June
PJ
Park
Hak Hee
HH
Kang
Tae Ryong
TR
Lee
D000943:Antigens, Differentiation / Q000032:analysis; D044383:Black People; D002122:Calcium Chloride / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division; D002469:Cell Separation / Q000379:methods*; D048430:Cell Shape; D002478:Cells, Cultured; D018920:Coculture Techniques; D000078404:Epidermal Cells; D018398:Homeodomain Proteins / Q000032:analysis; D006801:Humans; D007231:Infant, Newborn; D015603:Keratinocytes / Q000166:cytology* / Q000187:drug effects; D007633:Keratins / Q000032:analysis; D008544:Melanocytes / Q000166:cytology*; D051739:Microphthalmia-Associated Transcription Factor / Q000032:analysis; D009432:Neural Crest / Q000737:chemistry / Q000166:cytology* / Q000187:drug effects; D000072041:PAX3 Transcription Factor; D050809:POU Domain Factors / Q000032:analysis; D051761:Paired Box Transcription Factors / Q000032:analysis; D061251:Primary Cell Culture / Q000379:methods*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D044465:White People
D000943:Antigens, Differentiation; D018398:Homeodomain Proteins; C497234:MITF protein, human; D051739:Microphthalmia-Associated Transcription Factor; D000072041:PAX3 Transcription Factor; C489409:PAX3 protein, human; D050809:POU Domain Factors; D051761:Paired Box Transcription Factors; C085076:transcription factor Brn-2; D007633:Keratins; D002122:Calcium Chloride; D013755:Tetradecanoylphorbol Acetate
BRN2; calcium pulse; cell growth; human melanoblast; keratinocyte; melanocyte