Research Article

Laboratory Investigation (2009) 89, 844–856; doi:10.1038/labinvest.2009.48; published online 8 June 2009

Differential expression of stem-cell-associated markers in human hair follicle epithelial cells

Keita Inoue1, Noriyuki Aoi1, Takahiro Sato2, Yuji Yamauchi2, Hirotaka Suga1, Hitomi Eto1, Harunosuke Kato1, Jun Araki1 and Kotaro Yoshimura1

  1. 1Department of Plastic Surgery, University of Tokyo, Tokyo, Japan
  2. 2Division of Research and Development, Biomaster Inc., Yokohama, Japan

Correspondence: Professor K Yoshimura, MD, Department of Plastic Surgery, University of Tokyo School of Medicine, 7-3-1, Hongo, Bunkyo, Tokyo 113-8655, Japan. E-mail: yoshimura-pla@h.u-tokyo.ac.jp

Received 16 February 2009; Revised 31 March 2009; Accepted 7 April 2009; Published online 8 June 2009.

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Abstract

Several putative biomarkers have been suggested for identifying murine follicular stem cells; however, human hair follicles have a different pattern of biomarker expression, and follicular stem cell isolation methods have not been established. To isolate a stem cell population applicable to clinical settings, we conducted a comprehensive survey of the expression of stem-cell-associated (K15, CD200, CD34, and CD271) and other biomarkers (K1, K14, CD29, and CD49f) in immunohistological sections of the human epidermis and follicular outer root sheath (ORS). We also examined freshly isolated and cultured epidermal or follicular cells with single- and multicolor flow cytometry or immunocytochemistry. After sorting cells by CD200, CD34, and forward scatter (FSC) values (cell size), colony-forming assays were performed. We found that biomarkers were differentially expressed in the epidermis and ORS. Basal bulge cells were mainly K15+CD200+CD34CD271, and suprabasal cells were K15CD200+CD34CD271. We categorized follicular cells into nine subpopulations according to biomarker expression profiles. The CD200+CD34 bulge cells had much higher colony-forming abilities than the CD34+ population, and were divided into two subpopulations: a CD200+CD34FSChigh (K15-rich, basal) and a CD200+CD34FSClow (K15-poor, suprabasal) population. The former formed fewer but larger-sized colonies than the latter. Follicular epithelial cell cultivation resulted in loss of K15, CD200, CD34, and CD271 expression, but maintenance of K14, CD29, and CD49f expression. We found that the bulge contained two populations with different localizations, cell sizes, and colony-forming abilities. We showed that K15, CD200, CD34, and CD271 were useful biomarkers for characterizing freshly isolated human follicular epithelial cells in diverse stages of differentiation.

Keywords:

adult stem cells, colony formation, epidermis, flow cytometry, CD200, CD34

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