Histochemistry and Cell Biology

, Volume 133, Issue 4, pp 455–465

Differential expression of stem cell markers in human follicular bulge and interfollicular epidermal compartments

  • Shu Jiang
  • Longmei Zhao
  • Bhamini Purandare
  • Basil M. Hantash
Original Paper

Abstract

Although skin contains a number of stem cell repositories, their characterization has been hindered by a lack of specific markers and an unclear in vivo localization. In this study, we whole mounted single human scalp hair follicles and examined their profiles using in situ immunohistochemistry and multicolor immunofluorescence in search of markers to distinguish between stem cells residing in the interfollicular epidermis (IFE) and bulge. Our study revealed that expression of several biomarkers localized uniquely to the basal IFE (CD34 and CD117), bulge region (CD200), or both (CK15, CD49f, and CD29). In addition, we found that both basal IFE and bulge stem cells did not express CD71 or CD24 suggesting their potential utility as negative selection markers. Dermal papilla but not basal IFE or bulge stem cells expressed CD90, making it a potential positive selection marker for dermal hair follicle stem cells. The markers tested in this study may enable pursuit of cell sorting and purification strategies aimed at determining each stem cell population’s unique molecular signature.

Keywords

Epidermis Hair follicle Stem cell Cell surface marker 

Abbreviations

DP

Dermal papilla

HF

Hair follicle

IFE

Interfollicular epidermis

IRS

Inner root sheath

ORS

Outer root sheath

SG

Sebaceous glands

References

  1. Abbas O, Mahalingam M (2009) Epidermal stem cells: practical perspectives and potential uses. Br J Dermatol 161:228–236CrossRefPubMedGoogle Scholar
  2. Adams JC, Watt FM (1989) Fibronectin inhibits the terminal differentiation of human keratinocytes. Nature 340:307–309CrossRefPubMedGoogle Scholar
  3. Adams JC, Watt FM (1990) Changes in keratinocyte adhesion during terminal differentiation: reduction in fibronectin binding precedes alpha 5 beta 1 integrin loss from the cell surface. Cell 63:425–435CrossRefPubMedGoogle Scholar
  4. Akiyama M, Dale BA, Sun TT, Holbrook KA (1995) Characterization of hair follicle bulge in human fetal skin: the human fetal bulge is a pool of undifferentiated keratinocytes. J Invest Dermatol 105:844–850CrossRefPubMedGoogle Scholar
  5. Besmer P, Manova K, Duttlinger R, Huang EJ, Packer A, Gyssler C et al (1993) The kit-ligand (steel factor) and its receptor c-kit/W: pleiotropic roles in gametogenesis and melanogenesis. Dev Suppl 125–137Google Scholar
  6. Blanpain C, Lowry WE, Geoghegan A, Polak L, Fuchs E (2004) Self-renewal, multipotency, and the existence of two cell populations within an epithelial stem cell niche. Cell 118:635–648CrossRefPubMedGoogle Scholar
  7. Bosch P, Pratt SL, Stice SL (2006) Isolation, characterization, gene modification, and nuclear reprogramming of porcine mesenchymal stem cells. Biol Reprod 74:46–57CrossRefPubMedGoogle Scholar
  8. Botchkareva NV, Khlgatian M, Longley BJ, Botchkarev VA, Gilchrest BA (2001) SCF/c-kit signaling is required for cyclic regeneration of the hair pigmentation unit. FASEB J 15:645–658CrossRefPubMedGoogle Scholar
  9. Brakebusch C, Grose R, Quondamatteo F, Ramirez A, Jorcano JL, Pirro A et al (2000) Skin and hair follicle integrity is crucially dependent on beta 1 integrin expression on keratinocytes. EMBO J 19:3990–4003CrossRefPubMedGoogle Scholar
  10. Cotsarelis G (2006) Gene expression profiling gets to the root of human hair follicle stem cells. J Clin Invest 116:19–22CrossRefPubMedGoogle Scholar
  11. Cotsarelis G, Kaur P, Dhouailly D, Hengge U, Bickenbach J (1999) Epithelial stem cells in the skin: definition, markers, localization and functions. Exp Dermatol 8:80–88CrossRefPubMedGoogle Scholar
  12. Craig W, Kay R, Cutler RL, Lansdorp PM (1993) Expression of Thy-1 on human hematopoietic progenitor cells. J Exp Med 177:1331–1342CrossRefPubMedGoogle Scholar
  13. Florek M, Haase M, Marzesco AM, Freund D, Ehninger G, Huttner WB et al (2005) Prominin-1/CD133, a neural and hematopoietic stem cell marker, is expressed in adult human differentiated cells and certain types of kidney cancer. Cell Tissue Res 319:15–26CrossRefPubMedGoogle Scholar
  14. Ghazizadeh S, Taichman LB (2001) Multiple classes of stem cells in cutaneous epithelium: a lineage analysis of adult mouse skin. EMBO J 20:1215–1222CrossRefPubMedGoogle Scholar
  15. Ghazizadeh S, Taichman LB (2005) Organization of stem cells and their progeny in human epidermis. J Invest Dermatol 124:367–372CrossRefPubMedGoogle Scholar
  16. Giebel LB, Spritz RA (1991) Mutation of the KIT (mast/stem cell growth factor receptor) protooncogene in human piebaldism. Proc Natl Acad Sci USA 88:8696–8699CrossRefPubMedGoogle Scholar
  17. Grichnik JM, Ali WN, Burch JA, Byers JD, Garcia CA, Clark RE et al (1996) KIT expression reveals a population of precursor melanocytes in human skin. J Invest Dermatol 106:967–971CrossRefPubMedGoogle Scholar
  18. Hachiya A, Kobayashi A, Yoshida Y, Kitahara T, Takema Y, Imokawa G (2004) Biphasic expression of two paracrine melanogenic cytokines, stem cell factor and endothelin-1, in ultraviolet B-induced human melanogenesis. Am J Pathol 165:2099–2109PubMedGoogle Scholar
  19. Hentze MW, Kuhn LC (1996) Molecular control of vertebrate iron metabolism: mRNA-based regulatory circuits operated by iron, nitric oxide, and oxidative stress. Proc Natl Acad Sci USA 93:8175–8182CrossRefPubMedGoogle Scholar
  20. Hoang MP, Keady M, Mahalingam M (2009) Stem cell markers (cytokeratin 15, CD34 and nestin) in primary scarring and nonscarring alopecia. Br J Dermatol 160:609–615CrossRefPubMedGoogle Scholar
  21. Hotchin NA, Gandarillas A, Watt FM (1995) Regulation of cell surface beta 1 integrin levels during keratinocyte terminal differentiation. J Cell Biol 128:1209–1219CrossRefPubMedGoogle Scholar
  22. Inoue K, Aoi N, Sato T, Yamauchi Y, Suga H, Eto H et al (2009) Differential expression of stem-cell-associated markers in human hair follicle epithelial cells. Lab Invest 89:844–856CrossRefPubMedGoogle Scholar
  23. Ito M, Liu Y, Yang Z, Nguyen J, Liang F, Morris RJ et al (2005) Stem cells in the hair follicle bulge contribute to wound repair but not to homeostasis of the epidermis. Nat Med 11:1351–1354CrossRefPubMedGoogle Scholar
  24. Jahoda CA, Whitehouse J, Reynolds AJ, Hole N (2003) Hair follicle dermal cells differentiate into adipogenic and osteogenic lineages. Exp Dermatol 12:849–859CrossRefPubMedGoogle Scholar
  25. Janes SM, Lowell S, Hutter C (2002) Epidermal stem cells. J Pathol 197:479–491CrossRefPubMedGoogle Scholar
  26. Jih DM, Lyle S, Elenitsas R, Elder DE, Cotsarelis G (1999) Cytokeratin 15 expression in trichoepitheliomas and a subset of basal cell carcinomas suggests they originate from hair follicle stem cells. J Cutan Pathol 26:113–118CrossRefPubMedGoogle Scholar
  27. Kadmon G, Eckert M, Sammar M, Schachner M, Altevogt P (1992) Nectadrin, the heat-stable antigen, is a cell adhesion molecule. J Cell Biol 118:1245–1258CrossRefPubMedGoogle Scholar
  28. Kanitakis J, Bourchany D, Faure M, Claudy A (1999) Expression of the hair stem cell-specific keratin 15 in pilar tumors of the skin. Eur J Dermatol 9:363–365PubMedGoogle Scholar
  29. Kay R, Takei F, Humphries RK (1990) Expression cloning of a cDNA encoding M1/69-J11d heat-stable antigens. J Immunol 145:1952–1959PubMedGoogle Scholar
  30. Krause DS, Ito T, Fackler MJ, Smith OM, Collector MI, Sharkis SJ et al (1994) Characterization of murine CD34, a marker for hematopoietic progenitor and stem cells. Blood 84:691–701PubMedGoogle Scholar
  31. Lavker RM, Sun TT (1982) Heterogeneity in epidermal basal keratinocytes: morphological and functional correlations. Science 215:1239–1241CrossRefPubMedGoogle Scholar
  32. Lavker RM, Sun TT (2000) Epidermal stem cells: properties, markers, and location. Proc Natl Acad Sci USA 97:13473–13475CrossRefPubMedGoogle Scholar
  33. Levy V, Lindon C, Harfe BD, Morgan BA (2005) Distinct stem cell populations regenerate the follicle and interfollicular epidermis. Dev Cell 9:855–861CrossRefPubMedGoogle Scholar
  34. Levy V, Lindon C, Zheng Y, Harfe BD, Morgan BA (2007) Epidermal stem cells arise from the hair follicle after wounding. FASEB J 21:1358–1366CrossRefPubMedGoogle Scholar
  35. Li A, Kaur P (2005) FACS enrichment of human keratinocyte stem cells. Methods Mol Biol 289:87–96PubMedGoogle Scholar
  36. Li A, Simmons PJ, Kaur P (1998) Identification and isolation of candidate human keratinocyte stem cells based on cell surface phenotype. Proc Natl Acad Sci USA 95:3902–3907CrossRefPubMedGoogle Scholar
  37. Liu Y, Lyle S, Yang Z, Cotsarelis G (2003) Keratin 15 promoter targets putative epithelial stem cells in the hair follicle bulge. J Invest Dermatol 121:963–968CrossRefPubMedGoogle Scholar
  38. Lyle S, Christofidou-Solomidou M, Liu Y, Elder DE, Albelda S, Cotsarelis G (1998) The C8/144B monoclonal antibody recognizes cytokeratin 15 and defines the location of human hair follicle stem cells. J Cell Sci 111(Pt 21):3179–3188PubMedGoogle Scholar
  39. Ma DR, Yang EN, Lee ST (2004) A review: the location, molecular characterisation and multipotency of hair follicle epidermal stem cells. Ann Acad Med Singap 33:784–788PubMedGoogle Scholar
  40. Magnaldo T, Barrandon Y (1996) CD24 (heat stable antigen, nectadrin), a novel keratinocyte differentiation marker, is preferentially expressed in areas of the hair follicle containing the colony-forming cells. J Cell Sci 109(Pt 13):3035–3045PubMedGoogle Scholar
  41. Mason DY, Cordell JL, Gaulard P, Tse AG, Brown MH (1992) Immunohistological detection of human cytotoxic/suppressor T cells using antibodies to a CD8 peptide sequence. J Clin Pathol 45:1084–1088CrossRefPubMedGoogle Scholar
  42. Montagna W (1962) The structure and function of skin. Academic Press, New YorkGoogle Scholar
  43. Morris RJ, Liu Y, Marles L, Yang Z, Trempus C, Li S et al (2004) Capturing and profiling adult hair follicle stem cells. Nat Biotechnol 22:411–417CrossRefPubMedGoogle Scholar
  44. Morrison SJ, Weissman IL (1994) The long-term repopulating subset of hematopoietic stem cells is deterministic and isolatable by phenotype. Immunity 1:661–673CrossRefPubMedGoogle Scholar
  45. Nazareth MR, Broderick L, Simpson-Abelson MR, Kelleher RJ Jr, Yokota SJ, Bankert RB (2007) Characterization of human lung tumor-associated fibroblasts and their ability to modulate the activation of tumor-associated T cells. J Immunol 178:5552–5562PubMedGoogle Scholar
  46. Ohyama M, Terunuma A, Tock CL, Radonovich MF, Pise-Masison CA, Hopping SB et al (2006) Characterization and isolation of stem cell-enriched human hair follicle bulge cells. J Clin Invest 116:249–260CrossRefPubMedGoogle Scholar
  47. Oshima H, Rochat A, Kedzia C, Kobayashi K, Barrandon Y (2001) Morphogenesis and renewal of hair follicles from adult multipotent stem cells. Cell 104:233–245CrossRefPubMedGoogle Scholar
  48. Owens DM, Watt FM (2003) Contribution of stem cells and differentiated cells to epidermal tumours. Nat Rev Cancer 3:444–451CrossRefPubMedGoogle Scholar
  49. Poblet E, Jimenez F, Godinez JM, Pascual-Martin A, Izeta A (2006) The immunohistochemical expression of CD34 in human hair follicles: a comparative study with the bulge marker CK15. Clin Exp Dermatol 31:807–812CrossRefPubMedGoogle Scholar
  50. Raposio E, Guida C, Baldelli I, Curto M, Fiocca R, Kunkl A et al (2007) Characterization of multipotent cells from human adult hair follicles. Toxicol In Vitro 21:320–323CrossRefPubMedGoogle Scholar
  51. Reynolds AJ, Jahoda CA (1992) Cultured dermal papilla cells induce follicle formation and hair growth by transdifferentiation of an adult epidermis. Development 115:587–593PubMedGoogle Scholar
  52. Rosenblum MD, Olasz EB, Yancey KB, Woodliff JE, Lazarova Z, Gerber KA et al (2004) Expression of CD200 on epithelial cells of the murine hair follicle: a role in tissue-specific immune tolerance? J Invest Dermatol 123:880–887CrossRefPubMedGoogle Scholar
  53. Rosenblum MD, Yancey KB, Olasz EB, Truitt RL (2006) CD200, a “no danger” signal for hair follicles. J Dermatol Sci 41:165–174CrossRefPubMedGoogle Scholar
  54. Shmelkov SV, St Clair R, Lyden D, Rafii S (2005) AC133/CD133/Prominin-1. Int J Biochem Cell Biol 37:715–719CrossRefPubMedGoogle Scholar
  55. Stenn KS, Cotsarelis G (2005) Bioengineering the hair follicle: fringe benefits of stem cell technology. Curr Opin Biotechnol 16:493–497CrossRefPubMedGoogle Scholar
  56. Taylor G, Lehrer MS, Jensen PJ, Sun TT, Lavker RM (2000) Involvement of follicular stem cells in forming not only the follicle but also the epidermis. Cell 102:451–461CrossRefPubMedGoogle Scholar
  57. Trowbridge IS, Collawn JF, Hopkins CR (1993) Signal-dependent membrane protein trafficking in the endocytic pathway. Annu Rev Cell Biol 9:129–161CrossRefPubMedGoogle Scholar
  58. Tumbar T, Guasch G, Greco V, Blanpain C, Lowry WE, Rendl M et al (2004) Defining the epithelial stem cell niche in skin. Science 303:359–363CrossRefPubMedGoogle Scholar
  59. Waters JM, Richardson GD, Jahoda CA (2007) Hair follicle stem cells. Semin Cell Dev Biol 18:245–254CrossRefPubMedGoogle Scholar
  60. Watt FM, Hogan BL (2000) Out of Eden: stem cells and their niches. Science 287:1427–1430CrossRefPubMedGoogle Scholar
  61. Watt FM, Kubler MD, Hotchin NA, Nicholson LJ, Adams JC (1993) Regulation of keratinocyte terminal differentiation by integrin-extracellular matrix interactions. J Cell Sci 106(Pt 1):175–182PubMedGoogle Scholar
  62. Yin AH, Miraglia S, Zanjani ED, Almeida-Porada G, Ogawa M, Leary AG et al (1997) AC133, a novel marker for human hematopoietic stem and progenitor cells. Blood 90:5002–5012PubMedGoogle Scholar
  63. Yu K, Chen Z, Wang S, Gorczynski R (2005) Decreased alloreactivity using donor cells from mice expressing a CD200 transgene under control of a tetracycline-inducible promoter. Transplantation 80:394–401CrossRefPubMedGoogle Scholar
  64. Yu H, Fang D, Kumar SM, Li L, Nguyen TK, Acs G et al (2006) Isolation of a novel population of multipotent adult stem cells from human hair follicles. Am J Pathol 168:1879–1888CrossRefPubMedGoogle Scholar
  65. Zhu AJ, Haase I, Watt FM (1999) Signaling via beta1 integrins and mitogen-activated protein kinase determines human epidermal stem cell fate in vitro. Proc Natl Acad Sci USA 96:6728–6733CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Shu Jiang
    • 1
  • Longmei Zhao
    • 1
  • Bhamini Purandare
    • 2
  • Basil M. Hantash
    • 1
  1. 1.Division of Plastic Surgery, Department of SurgeryStanford University School of MedicineStanfordUSA
  2. 2.San Jose State UniversitySan JoseUSA

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