In situ localization of epidermal stem cells using a novel multi epitope ligand cartography approach.

Precise knowledge of the frequency and localization of epidermal stem cells within skin tissue would further our understanding of their role in maintaining skin homeostasis. As a novel approach we used the recently developed method of multi epitope ligand cartography, applying a set of described putative epidermal stem cell markers. Bioinformatic evaluation of the data led to the identification of several discrete basal keratinocyte populations, but none of them displayed the complete stem cell marker set. The distribution of the keratinocyte populations within the tissue was remarkably heterogeneous, but determination of distance relationships revealed a population of quiescent cells highly expressing p63 and the integrins alpha(6)/beta(1) that represent origins of a gradual differentiation lineage. This population comprises about 6% of all basal cells, shows a scattered distribution pattern and could also be found in keratinocyte holoclone colonies. The data suggest that this population identifies interfollicular epidermal stem cells.

[1]  F. Watt,et al.  beta-catenin signalling modulates proliferative potential of human epidermal keratinocytes independently of intercellular adhesion. , 1999, Development.

[2]  P. Kaur.  Interfollicular epidermal stem cells: identification, challenges, potential. , 2006, The Journal of investigative dermatology.

[3]  R. Ghadially,et al.  Measuring stem cell frequency in epidermis: A quantitative in vivo functional assay for long-term repopulating cells , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[4]  Y. Barrandon,et al.  Three clonal types of keratinocyte with different capacities for multiplication. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[5]  D. Ponzin,et al.  p63 identifies keratinocyte stem cells , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[6]  F. Watt,et al.  Stem cell patterning and fate in human epidermis , 1995, Cell.

[7]  W. Fitzgerald,et al.  Isolation and Clonal Analysis of Human Epidermal Keratinocyte Stem Cells in Long‐Term Culture , 2003, Stem cells.

[8]  C. Potten.  THE EPIDERMAL PROLIFERATIVE UNIT: THE POSSIBLE ROLE OF THE CENTRAL BASAL CELL , 1974, Cell and tissue kinetics.

[9]  Fiona M. Watt,et al.  Separation of human epidermal stem cells from transit amplifying cells on the basis of differences in integrin function and expression , 1993, Cell.

[10]  Amy Li,et al.  Identification and isolation of candidate human keratinocyte stem cells based on cell surface phenotype. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[11]  F M Watt,et al.  The spatial relationship between stem cells and their progeny in the basal layer of human epidermis: a new view based on whole-mount labelling and lineage analysis. , 1999, Development.

[12]  Jiaxi Zhou,et al.  Enrichment of putative human epidermal stem cells based on cell size and collagen type IV adhesiveness , 2008, Cell Research.

[13]  J. Cairns,et al.  The segregation of DNA in epithelial stem cells , 1978, Cell.

[14]  A. Yang,et al.  p63, a p53 homolog at 3q27-29, encodes multiple products with transactivating, death-inducing, and dominant-negative activities. , 1998, Molecular cell.

[15]  B. Nickoloff,et al.  Estimation of size of clonal unit for keratinocytes in normal human skin. , 2002, Archives of pathology & laboratory medicine.

[16]  Benjamin D. Simons,et al.  A single type of progenitor cell maintains normal epidermis , 2007, Nature.

[17]  F. Pontén,et al.  Mosaic pattern of maternal and paternal keratinocyte clones in normal human epidermis revealed by analysis of X-chromosome inactivation. , 2001, The Journal of investigative dermatology.

[18]  G. Weinstein,et al.  Cell proliferation in normal epidermis. , 1984, The Journal of investigative dermatology.

[19]  S. Olsen,et al.  Identification of a cell surface protein with a role in stimulating human keratinocyte proliferation, expressed during development and carcinogenesis. , 1997, The Journal of investigative dermatology.

[20]  F. Watt,et al.  Role of melanoma chondroitin sulphate proteoglycan in patterning stem cells in human interfollicular epidermis , 2003, Development.

[21]  H. Cho,et al.  Isolation of human epidermal stem cells by adherence and the reconstruction of skin equivalents , 2004, Cellular and Molecular Life Sciences CMLS.

[22]  F. McKeon,et al.  p63 Is Essential for the Proliferative Potential of Stem Cells in Stratified Epithelia , 2007, Cell.

[23]  S. Ghazizadeh,et al.  Regeneration of multilineage skin epithelia by differentiated keratinocytes. , 2010, The Journal of investigative dermatology.

[24]  Walter Schubert,et al.  A three‐symbol code for organized proteomes based on cyclical imaging of protein locations , 2007, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[25]  J. Bickenbach.  Identification and behavior of label-retaining cells in oral mucosa and skin. , 1981, Journal of dental research.

[26]  W. Schubert,et al.  Analyzing proteome topology and function by automated multidimensional fluorescence microscopy , 2006, Nature Biotechnology.

[27]  M. Grever,et al.  Predictive statistics and artificial intelligence in the U.S. National Cancer Institute's drug discovery program for cancer and AIDS , 1994, Stem cells.

[28]  Amy Li,et al.  Side population in adult murine epidermis exhibits phenotypic and functional characteristics of keratinocyte stem cells , 2006, Proceedings of the National Academy of Sciences.