Generation of human melanocytes from induced pluripotent stem cells

Shigeki Ota, Yoichi Imaizumi, Yohei Okada, Wado Akamatsu, Reiko Kuwahara, Manabu Ohyama, Masayuki Amagai, Yumi Matsuzaki, Shinya Yamanaka, Hideyuki Okano, Yutaka Kawakami

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

Epidermal melanocytes play an important role in protecting the skin from UV rays, and their functional impairment results in pigment disorders. Additionally, melanomas are considered to arise from mutations that accumulate in melanocyte stem cells. The mechanisms underlying melanocyte differentiation and the defining characteristics of melanocyte stem cells in humans are, however, largely unknown. In the present study, we set out to generate melanocytes from human iPS cells in vitro, leading to a preliminary investigation of the mechanisms of human melanocyte differentiation. We generated iPS cell lines from human dermal fibroblasts using the Yamanaka factors (SOX2, OCT3/4, and KLF4, with or without c-MYC). These iPS cell lines were subsequently used to form embryoid bodies (EBs) and then differentiated into melanocytes via culture supplementation with Wnt3a, SCF, and ET-3. Seven weeks after inducing differentiation, pigmented cells expressing melanocyte markers such as MITF, tyrosinase, SILV, and TYRP1, were detected. Melanosomes were identified in these pigmented cells by electron microscopy, and global gene expression profiling of the pigmented cells showed a high similarity to that of human primary foreskin-derived melanocytes, suggesting the successful generation of melanocytes from iPS cells. This in vitro differentiation system should prove useful for understanding human melanocyte biology and revealing the mechanism of various pigment cell disorders, including melanoma.

Original languageEnglish
Article numbere16182
JournalPLoS One
Volume6
Issue number1
DOIs
Publication statusPublished - 2011

Fingerprint

Induced Pluripotent Stem Cells
melanocytes
Melanocytes
Stem cells
Cells
Pigments
Monophenol Monooxygenase
Fibroblasts
Gene expression
Electron microscopy
Skin
melanoma
stem cells
Melanoma
induced pluripotent stem cells
Stem Cells
pigments
cell lines
cells
Embryoid Bodies

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Ota, S., Imaizumi, Y., Okada, Y., Akamatsu, W., Kuwahara, R., Ohyama, M., ... Kawakami, Y. (2011). Generation of human melanocytes from induced pluripotent stem cells. PLoS One, 6(1), [e16182]. https://doi.org/10.1371/journal.pone.0016182

Generation of human melanocytes from induced pluripotent stem cells. / Ota, Shigeki; Imaizumi, Yoichi; Okada, Yohei; Akamatsu, Wado; Kuwahara, Reiko; Ohyama, Manabu; Amagai, Masayuki; Matsuzaki, Yumi; Yamanaka, Shinya; Okano, Hideyuki; Kawakami, Yutaka.

In: PLoS One, Vol. 6, No. 1, e16182, 2011.

Research output: Contribution to journalArticle

Ota, S, Imaizumi, Y, Okada, Y, Akamatsu, W, Kuwahara, R, Ohyama, M, Amagai, M, Matsuzaki, Y, Yamanaka, S, Okano, H & Kawakami, Y 2011, 'Generation of human melanocytes from induced pluripotent stem cells', PLoS One, vol. 6, no. 1, e16182. https://doi.org/10.1371/journal.pone.0016182
Ota S, Imaizumi Y, Okada Y, Akamatsu W, Kuwahara R, Ohyama M et al. Generation of human melanocytes from induced pluripotent stem cells. PLoS One. 2011;6(1). e16182. https://doi.org/10.1371/journal.pone.0016182
Ota, Shigeki ; Imaizumi, Yoichi ; Okada, Yohei ; Akamatsu, Wado ; Kuwahara, Reiko ; Ohyama, Manabu ; Amagai, Masayuki ; Matsuzaki, Yumi ; Yamanaka, Shinya ; Okano, Hideyuki ; Kawakami, Yutaka. / Generation of human melanocytes from induced pluripotent stem cells. In: PLoS One. 2011 ; Vol. 6, No. 1.
@article{4d7b29d8d03a45c6b5f0d25d5e8a6cfc,
title = "Generation of human melanocytes from induced pluripotent stem cells",
abstract = "Epidermal melanocytes play an important role in protecting the skin from UV rays, and their functional impairment results in pigment disorders. Additionally, melanomas are considered to arise from mutations that accumulate in melanocyte stem cells. The mechanisms underlying melanocyte differentiation and the defining characteristics of melanocyte stem cells in humans are, however, largely unknown. In the present study, we set out to generate melanocytes from human iPS cells in vitro, leading to a preliminary investigation of the mechanisms of human melanocyte differentiation. We generated iPS cell lines from human dermal fibroblasts using the Yamanaka factors (SOX2, OCT3/4, and KLF4, with or without c-MYC). These iPS cell lines were subsequently used to form embryoid bodies (EBs) and then differentiated into melanocytes via culture supplementation with Wnt3a, SCF, and ET-3. Seven weeks after inducing differentiation, pigmented cells expressing melanocyte markers such as MITF, tyrosinase, SILV, and TYRP1, were detected. Melanosomes were identified in these pigmented cells by electron microscopy, and global gene expression profiling of the pigmented cells showed a high similarity to that of human primary foreskin-derived melanocytes, suggesting the successful generation of melanocytes from iPS cells. This in vitro differentiation system should prove useful for understanding human melanocyte biology and revealing the mechanism of various pigment cell disorders, including melanoma.",
author = "Shigeki Ota and Yoichi Imaizumi and Yohei Okada and Wado Akamatsu and Reiko Kuwahara and Manabu Ohyama and Masayuki Amagai and Yumi Matsuzaki and Shinya Yamanaka and Hideyuki Okano and Yutaka Kawakami",
year = "2011",
doi = "10.1371/journal.pone.0016182",
language = "English",
volume = "6",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "1",

}

TY - JOUR

T1 - Generation of human melanocytes from induced pluripotent stem cells

AU - Ota, Shigeki

AU - Imaizumi, Yoichi

AU - Okada, Yohei

AU - Akamatsu, Wado

AU - Kuwahara, Reiko

AU - Ohyama, Manabu

AU - Amagai, Masayuki

AU - Matsuzaki, Yumi

AU - Yamanaka, Shinya

AU - Okano, Hideyuki

AU - Kawakami, Yutaka

PY - 2011

Y1 - 2011

N2 - Epidermal melanocytes play an important role in protecting the skin from UV rays, and their functional impairment results in pigment disorders. Additionally, melanomas are considered to arise from mutations that accumulate in melanocyte stem cells. The mechanisms underlying melanocyte differentiation and the defining characteristics of melanocyte stem cells in humans are, however, largely unknown. In the present study, we set out to generate melanocytes from human iPS cells in vitro, leading to a preliminary investigation of the mechanisms of human melanocyte differentiation. We generated iPS cell lines from human dermal fibroblasts using the Yamanaka factors (SOX2, OCT3/4, and KLF4, with or without c-MYC). These iPS cell lines were subsequently used to form embryoid bodies (EBs) and then differentiated into melanocytes via culture supplementation with Wnt3a, SCF, and ET-3. Seven weeks after inducing differentiation, pigmented cells expressing melanocyte markers such as MITF, tyrosinase, SILV, and TYRP1, were detected. Melanosomes were identified in these pigmented cells by electron microscopy, and global gene expression profiling of the pigmented cells showed a high similarity to that of human primary foreskin-derived melanocytes, suggesting the successful generation of melanocytes from iPS cells. This in vitro differentiation system should prove useful for understanding human melanocyte biology and revealing the mechanism of various pigment cell disorders, including melanoma.

AB - Epidermal melanocytes play an important role in protecting the skin from UV rays, and their functional impairment results in pigment disorders. Additionally, melanomas are considered to arise from mutations that accumulate in melanocyte stem cells. The mechanisms underlying melanocyte differentiation and the defining characteristics of melanocyte stem cells in humans are, however, largely unknown. In the present study, we set out to generate melanocytes from human iPS cells in vitro, leading to a preliminary investigation of the mechanisms of human melanocyte differentiation. We generated iPS cell lines from human dermal fibroblasts using the Yamanaka factors (SOX2, OCT3/4, and KLF4, with or without c-MYC). These iPS cell lines were subsequently used to form embryoid bodies (EBs) and then differentiated into melanocytes via culture supplementation with Wnt3a, SCF, and ET-3. Seven weeks after inducing differentiation, pigmented cells expressing melanocyte markers such as MITF, tyrosinase, SILV, and TYRP1, were detected. Melanosomes were identified in these pigmented cells by electron microscopy, and global gene expression profiling of the pigmented cells showed a high similarity to that of human primary foreskin-derived melanocytes, suggesting the successful generation of melanocytes from iPS cells. This in vitro differentiation system should prove useful for understanding human melanocyte biology and revealing the mechanism of various pigment cell disorders, including melanoma.

UR - http://www.scopus.com/inward/record.url?scp=79251578074&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79251578074&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0016182

DO - 10.1371/journal.pone.0016182

M3 - Article

C2 - 21249204

AN - SCOPUS:79251578074

VL - 6

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 1

M1 - e16182

ER -