Characterization and isolation of stem cell–enriched human hair follicle bulge cells
Manabu Ohyama, … , Mark C. Udey, Jonathan C. Vogel
Manabu Ohyama, … , Mark C. Udey, Jonathan C. Vogel
Published January 4, 2006
Citation Information: J Clin Invest. 2006;116(1):249-260. https://doi.org/10.1172/JCI26043.
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Research Article Dermatology

Characterization and isolation of stem cell–enriched human hair follicle bulge cells

Abstract

The human hair follicle bulge is an important niche for keratinocyte stem cells (KSCs). Elucidation of human bulge cell biology could be facilitated by analysis of global gene expression profiles and identification of unique cell-surface markers. The lack of distinctive bulge morphology in human hair follicles has hampered studies of bulge cells and KSCs. In this study, we determined the distribution of label-retaining cells to define the human anagen bulge. Using navigated laser capture microdissection, bulge cells and outer root sheath cells from other follicle regions were obtained and analyzed with cDNA microarrays. Gene transcripts encoding inhibitors of WNT and activin/bone morphogenic protein signaling were overrepresented in the bulge, while genes responsible for cell proliferation were underrepresented, consistent with the existence of quiescent noncycling KSCs in anagen follicles. Positive markers for bulge cells included CD200, PHLDA1, follistatin, and frizzled homolog 1, while CD24, CD34, CD71, and CD146 were preferentially expressed by non-bulge keratinocytes. Importantly, CD200+ cells (CD200hiCD24loCD34loCD71loCD146lo) obtained from hair follicle suspensions demonstrated high colony-forming efficiency in clonogenic assays, indicating successful enrichment of living human bulge stem cells. The stem cell behavior of enriched bulge cells and their utility for gene therapy and hair regeneration will need to be assessed in in vivo assays.

Authors

Manabu Ohyama, Atsushi Terunuma, Christine L. Tock, Michael F. Radonovich, Cynthia A. Pise-Masison, Steven B. Hopping, John N. Brady, Mark C. Udey, Jonathan C. Vogel

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Figure 5

CD59 and CD200 are expressed in human anagen hair follicle, and CD200 is a positive cell-surface marker for the bulge ORS cells.

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CD59 and CD200 are expressed in human anagen hair follicle, and CD200 is...
(A) Immunohistochemically, CD59 was globally expressed at all ORS levels (vertical section) and upregulated in the bulge ORS (transverse sections). Scale bars: 50 μm. In FACS analysis, most mid-follicle cells were CD59+, and FSThi bulge cells were CD59+ (CD59 staining prior to fixation). (B) CD200 was preferentially expressed on the defined bulge ORS and the companion layer of human anagen hair follicles. Scale bars: 50 μm. By FACS analysis, CD200 was detectable on the mid-follicle cells, and approximately 40% of CD200hi cells were FST-double-positive bulge cells (CD200 staining before fixation).