Restoration of the intrinsic properties of human dermal papilla in vitro

Manabu Ohyama, Tetsuro Kobayashi, Takashi Sasaki, Atsushi Shimizu, Masayuki Amagai

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

The dermal papilla (DP) plays pivotal roles in hair follicle morphogenesis and cycling. However, characterization and/or propagation of human DPs have been unsatisfactory because of the lack of efficient isolation methods and the loss of innate characteristics in vitro. We hypothesized that culture conditions sustaining the intrinsic molecular signature of the human DP could facilitate expansion of functional DP cells. To test this, we first characterized the global gene expression profile of microdissected, non-cultured human DPs. We performed a 'two-step' microarray analysis to exclude the influence of unwanted contaminants in isolated DPs and successfully identified 118 human DP signature genes, including 38 genes listed in the mouse DP signature. The bioinformatics analysis of the DP gene list revealed that WNT, BMP and FGF signaling pathways were upregulated in intact DPs and addition of 6-bromoindirubin-39-oxime, recombinant BMP2 and basic FGF to stimulate these respective signaling pathways resulted in maintained expression of in situ DP signature genes in primarily cultured human DP cells. More importantly, the exposure to these stimulants restored normally reduced DP biomarker expression in conventionally cultured DP cells. Cell growth was moderate in the newly developed culture medium. However, rapid DP cell expansion by conventional culture followed by the restoration by defined activators provided a sufficient number of DP cells that demonstrated characteristic DP activities in functional assays. The study reported here revealed previously unreported molecular mechanisms contributing to human DP properties and describes a useful technique for the investigation of human DP biology and hair follicle bioengineering.

Original languageEnglish
Pages (from-to)4114-4125
Number of pages12
JournalJournal of Cell Science
Volume125
Issue number17
DOIs
Publication statusPublished - 2012 Sep 1

Fingerprint

Skin
Hair Follicle
In Vitro Techniques
Genes
Bioengineering
Oximes
Microarray Analysis
Computational Biology
Morphogenesis
Transcriptome
Culture Media
Biomarkers
Growth

Keywords

  • Cell culture
  • Dermal papilla
  • Human
  • Molecular signature

ASJC Scopus subject areas

  • Cell Biology

Cite this

Restoration of the intrinsic properties of human dermal papilla in vitro. / Ohyama, Manabu; Kobayashi, Tetsuro; Sasaki, Takashi; Shimizu, Atsushi; Amagai, Masayuki.

In: Journal of Cell Science, Vol. 125, No. 17, 01.09.2012, p. 4114-4125.

Research output: Contribution to journalArticle

Ohyama, Manabu ; Kobayashi, Tetsuro ; Sasaki, Takashi ; Shimizu, Atsushi ; Amagai, Masayuki. / Restoration of the intrinsic properties of human dermal papilla in vitro. In: Journal of Cell Science. 2012 ; Vol. 125, No. 17. pp. 4114-4125.
@article{b8dc936d83274f759e46e484815d05c7,
title = "Restoration of the intrinsic properties of human dermal papilla in vitro",
abstract = "The dermal papilla (DP) plays pivotal roles in hair follicle morphogenesis and cycling. However, characterization and/or propagation of human DPs have been unsatisfactory because of the lack of efficient isolation methods and the loss of innate characteristics in vitro. We hypothesized that culture conditions sustaining the intrinsic molecular signature of the human DP could facilitate expansion of functional DP cells. To test this, we first characterized the global gene expression profile of microdissected, non-cultured human DPs. We performed a 'two-step' microarray analysis to exclude the influence of unwanted contaminants in isolated DPs and successfully identified 118 human DP signature genes, including 38 genes listed in the mouse DP signature. The bioinformatics analysis of the DP gene list revealed that WNT, BMP and FGF signaling pathways were upregulated in intact DPs and addition of 6-bromoindirubin-39-oxime, recombinant BMP2 and basic FGF to stimulate these respective signaling pathways resulted in maintained expression of in situ DP signature genes in primarily cultured human DP cells. More importantly, the exposure to these stimulants restored normally reduced DP biomarker expression in conventionally cultured DP cells. Cell growth was moderate in the newly developed culture medium. However, rapid DP cell expansion by conventional culture followed by the restoration by defined activators provided a sufficient number of DP cells that demonstrated characteristic DP activities in functional assays. The study reported here revealed previously unreported molecular mechanisms contributing to human DP properties and describes a useful technique for the investigation of human DP biology and hair follicle bioengineering.",
keywords = "Cell culture, Dermal papilla, Human, Molecular signature",
author = "Manabu Ohyama and Tetsuro Kobayashi and Takashi Sasaki and Atsushi Shimizu and Masayuki Amagai",
year = "2012",
month = "9",
day = "1",
doi = "10.1242/jcs.105700",
language = "English",
volume = "125",
pages = "4114--4125",
journal = "Journal of Cell Science",
issn = "0021-9533",
publisher = "Company of Biologists Ltd",
number = "17",

}

TY - JOUR

T1 - Restoration of the intrinsic properties of human dermal papilla in vitro

AU - Ohyama, Manabu

AU - Kobayashi, Tetsuro

AU - Sasaki, Takashi

AU - Shimizu, Atsushi

AU - Amagai, Masayuki

PY - 2012/9/1

Y1 - 2012/9/1

N2 - The dermal papilla (DP) plays pivotal roles in hair follicle morphogenesis and cycling. However, characterization and/or propagation of human DPs have been unsatisfactory because of the lack of efficient isolation methods and the loss of innate characteristics in vitro. We hypothesized that culture conditions sustaining the intrinsic molecular signature of the human DP could facilitate expansion of functional DP cells. To test this, we first characterized the global gene expression profile of microdissected, non-cultured human DPs. We performed a 'two-step' microarray analysis to exclude the influence of unwanted contaminants in isolated DPs and successfully identified 118 human DP signature genes, including 38 genes listed in the mouse DP signature. The bioinformatics analysis of the DP gene list revealed that WNT, BMP and FGF signaling pathways were upregulated in intact DPs and addition of 6-bromoindirubin-39-oxime, recombinant BMP2 and basic FGF to stimulate these respective signaling pathways resulted in maintained expression of in situ DP signature genes in primarily cultured human DP cells. More importantly, the exposure to these stimulants restored normally reduced DP biomarker expression in conventionally cultured DP cells. Cell growth was moderate in the newly developed culture medium. However, rapid DP cell expansion by conventional culture followed by the restoration by defined activators provided a sufficient number of DP cells that demonstrated characteristic DP activities in functional assays. The study reported here revealed previously unreported molecular mechanisms contributing to human DP properties and describes a useful technique for the investigation of human DP biology and hair follicle bioengineering.

AB - The dermal papilla (DP) plays pivotal roles in hair follicle morphogenesis and cycling. However, characterization and/or propagation of human DPs have been unsatisfactory because of the lack of efficient isolation methods and the loss of innate characteristics in vitro. We hypothesized that culture conditions sustaining the intrinsic molecular signature of the human DP could facilitate expansion of functional DP cells. To test this, we first characterized the global gene expression profile of microdissected, non-cultured human DPs. We performed a 'two-step' microarray analysis to exclude the influence of unwanted contaminants in isolated DPs and successfully identified 118 human DP signature genes, including 38 genes listed in the mouse DP signature. The bioinformatics analysis of the DP gene list revealed that WNT, BMP and FGF signaling pathways were upregulated in intact DPs and addition of 6-bromoindirubin-39-oxime, recombinant BMP2 and basic FGF to stimulate these respective signaling pathways resulted in maintained expression of in situ DP signature genes in primarily cultured human DP cells. More importantly, the exposure to these stimulants restored normally reduced DP biomarker expression in conventionally cultured DP cells. Cell growth was moderate in the newly developed culture medium. However, rapid DP cell expansion by conventional culture followed by the restoration by defined activators provided a sufficient number of DP cells that demonstrated characteristic DP activities in functional assays. The study reported here revealed previously unreported molecular mechanisms contributing to human DP properties and describes a useful technique for the investigation of human DP biology and hair follicle bioengineering.

KW - Cell culture

KW - Dermal papilla

KW - Human

KW - Molecular signature

UR - http://www.scopus.com/inward/record.url?scp=84868210284&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84868210284&partnerID=8YFLogxK

U2 - 10.1242/jcs.105700

DO - 10.1242/jcs.105700

M3 - Article

VL - 125

SP - 4114

EP - 4125

JO - Journal of Cell Science

JF - Journal of Cell Science

SN - 0021-9533

IS - 17

ER -