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First published online March 30, 2006
Stem Cells Vol. 24 No. 7 July 2006, pp. 1668 -1677
doi:10.1634/stemcells.2005-0414; www.StemCells.com
© 2006 AlphaMed Press

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EMBRYONIC STEM CELLS

Defining the Conditions for the Generation of Melanocytes from Human Embryonic Stem Cells

Dong Fanga, Kim Leisheara, Thiennga K. Nguyena, Rena Finkoa, Kun Caia, Mizuho Fukunagaa, Ling Lia, Patricia A. Brafforda, Angela N. Kulpa, Xiaowei Xub, Keiran S. M. Smalleya, Meenhard Herlyna

aProgram of Molecular and Cellular Oncogenesis, The Wistar Institute, Philadelphia, Pennsylvania, USA;
bDepartment of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, USA

Key Words. Human embryonic stem cells • Melanocytes • Development • Wnt3a • Stem cell factor • Endothelin-3

Correspondence: Meenhard Herlyn, D.V.M., D.S.C.,Program of Molecular and Cellular Oncogenesis, The Wistar Institute, Philadelphia, Pennsylvania 19104, USA.Telephone: 215-898-3950;Fax: 215-898-0980;email: herlynm{at}wistar.org

Received August 26, 2005; accepted for publication March 22, 2006.
First published online in STEM CELLS EXPRESS   March 30, 2006.


Because of their undifferentiated nature, human embryonic stem cells (hESCs) are an ideal model system for studying both normal human development and the processes that underlie disease. In the current study, we describe an efficient method for differentiating hESCs into a melanocyte population within 4–6 weeks using three growth factors: Wnt3a, endothelin-3, and stem cell factor. The hESC-derived melanocytes expressed melanocyte markers (such as microphthalmia-associated transcription factor and tyrosinase), developed melanosomes, and produced melanin. They retained the melanocyte phenotype during long-term cell culture (>90 days) and, when incorporated into human reconstructed skin, homed to the appropriate location along the basement membrane in the same manner as epidermis-derived melanocytes. They maintained a stable phenotype even after grafting of the reconstructs to immunodeficient mice. Over time in culture, the hESC-derived melanocytes lost expression of telomerase and underwent senescence. In summary, we have shown for the first time the differentiation of hESCs into melanocytes. This method provides a novel in vitro system for studying the development biology of human melanocytes.




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