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Vol. 22, No. 2, 2009
Issue release date: February 2009
Skin Pharmacol Physiol 2009;22:55–62

Stem Cells and Tissue-Engineered Skin

Charruyer A. · Ghadially R.
Department of Dermatology, University of California and Veterans Affairs Medical Center, San Francisco, Calif., USA

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Advances in tissue engineering of skin are needed for clinical applications (as in wound healing and gene therapy) for cutaneous and systemic diseases. In this paper we review the use of epidermal stem cells as a source of cells to improve tissue-engineered skin. We discuss the importance and limitations of epidermal stem cell isolation using biomarkers, in quest of a pure stem cell preparation, as well as the culture conditions necessary to maintain this purity as required for a qualitatively superior and long-lasting engineered skin. Finally, we review the advantages of using additional multipotent stem cell sources to functionally and cosmetically optimize the engineered tissue.

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  1. Meuli M, Raghunath M: Burns. Part 2. Tops and flops using cultured epithelial autografts in children. Pediatr Surg Int 1997;12:471–477.
  2. Bianco P, Robey PG: Stem cells in tissue engineering. Nature 2001;414:118–121.
  3. Dunnwald M, Tomanek-Chalkley A, Alexandrunas D, Fishbaugh J, Bickenbach JR: Isolating a pure population of epidermal stem cells for use in tissue engineering. Exp Dermatol 2001;10:45–54.
  4. Pellegrini G, Ranno R, Stracuzzi G, Bondanza S, Guerra L, Zambruno G, Micali G, De Luca M: The control of epidermal stem cells (holoclones) in the treatment of massive full-thickness burns with autologous keratinocytes cultured on fibrin. Transplantation 1999;68:868–879.
  5. Ito M, Liu Y, Yang Z, Nguyen J, Liang F, Morris RJ, Cotsarelis G: Stem cells in the hair follicle bulge contribute to wound repair but not to homeostasis of the epidermis. Nat Med 2005;11:1351–1354.
  6. Langton AK, Herrick SE, Headon DJ: An extended epidermal response heals cutaneous wounds in the absence of a hair follicle stem cell contribution. J Invest Dermatol 2008;128:1311–1318.
  7. Levy V, Lindon C, Zheng Y, Harfe BD, Morgan BA: Epidermal stem cells arise from the hair follicle after wounding. FASEB J 2007;21:1358–1366.
  8. Uchida N, Fleming WH, Alpern EJ, Weissman IL: Heterogeneity of hematopoietic stem cells. Curr Opin Immunol 1993;5:177–184.
  9. Li A, Simmons PJ, Kaur P: Identification and isolation of candidate human keratinocyte stem cells based on cell surface phenotype. Proc Natl Acad Sci USA 1998;95:3902–3907.
  10. Terunuma A, Kapoor V, Yee C, Telford WG, Udey MC, Vogel JC: Stem cell activity of human side population and α6 integrin-bright keratinocytes defined by a quantitative in vivo assay. Stem Cells 2007;25:664–669.
  11. Bickenbach JR, Chism E: Selection and extended growth of murine epidermal stem cells in culture. Exp Cell Res 1998;244:184–195.
  12. Strachan LR, Scalapino KJ, Lawrence HJ, Ghadially R: Rapid adhesion to collagen isolates murine keratinocytes with limited long-term repopulating ability in vivo despite high clonogenicity in vitro. Stem Cells 2008;26:235–243.
  13. Bickenbach JR: Identification and behavior of label-retaining cells in oral mucosa and skin. J Dent Res 1981;60 :1611–1620.
  14. Kiel MJ, He S, Ashkenazi R, Gentry SN, Teta M, Kushner JA, Jackson TL, Morrison SJ: Haematopoietic stem cells do not asymmetrically segregate chromosomes or retain BrdU. Nature 2007;449:238–242.
  15. Morris RJ, Fischer SM, Klein-Szanto AJ, Slaga TJ: Subpopulations of primary adult murine epidermal basal cells sedimented on density gradients. Cell Tissue Kinet 1990;23:587–602.
  16. Terunuma A, Jackson KL, Kapoor V, Telford WG, Vogel JC: Side population keratinocytes resembling bone marrow side population stem cells are distinct from label-retaining keratinocyte stem cells. J Invest Dermatol 2003;121:1095–1103.
  17. Triel C, Vestergaard ME, Bolund L, Jensen TG, Jensen UB: Side population cells in human and mouse epidermis lack stem cell characteristics. Exp Cell Res 2004;295:79–90.
  18. Tani H, Morris RJ, Kaur P: Enrichment for murine keratinocyte stem cells based on cell surface phenotype. Proc Natl Acad Sci USA 2000;97:10960–10965.
  19. Kim DS, Cho HJ, Choi HR, Kwon SB, Park KC: Isolation of human epidermal stem cells by adherence and the reconstruction of skin equivalents. Cell Mol Life Sci 2004;61:2774–2781.
  20. Watt FM: Studies with cultured human epidermal keratinocytes: potential relevance to corneal wound healing. Eye 1994;8:161–162.
  21. Jones PH: Isolation and characterization of human epidermal stem cells. Clin Sci (Lond) 1996;91:141–146.
  22. Jones PH, Harper S, Watt FM: Stem cell patterning and fate in human epidermis. Cell 1995;80:83–93.
  23. Brakebusch C, Grose R, Quondamatteo F, Ramirez A, Jorcano JL, Pirro A, Svensson M, Herken R, Sasaki T, Timpl R, Werner S, Fässler R: Skin and hair follicle integrity is crucially dependent on β1 integrin expression on keratinocytes. EMBO J 2000;19:3990–4003.
  24. Raghavan S, Bauer C, Mundschau G, Li Q, Fuchs E: Conditional ablation of beta1 integrin in skin. Severe defects in epidermal proliferation, basement membrane formation, and hair follicle invagination. J Cell Biol 2000;150:1149–1160.
  25. Potten CS: The epidermal proliferative unit: the possible role of the central basal cell. Cell Tissue Kinet 1974;7:77–88.
  26. Rowe L, Dixon WJ: Clustering and control of mitotic activity in human epidermis. J Invest Dermatol 1972;58:16–23.
  27. Potten CS, Morris RJ: Epithelial stem cells in vivo. J Cell Sci Suppl 1988;10:45–62.
  28. Braun KM, Watt FM: Epidermal label-retaining cells: background and recent applications. J Investig Dermatol Symp Proc 2004;9:196–201.
  29. Lin KK, Goodell M A: Purification of hematopoietic stem cells using the side population. Methods Enzymol 2006;420:255–264.
  30. Uezumi, A, Ojima K, Fukada S, Ikemoto M, Masuda S, Miyagoe-Suzuki Y, Takeda S: Functional heterogeneity of side population cells in skeletal muscle. Biochem Biophys Res Commun 2006;341:864–873.
  31. Kohno K, Sato S, Takano H, Matsuo K, Kuwano M: The direct activation of human multidrug resistance gene (MDR1) by anticancer agents. Biochem Biophys Res Commun 1989;165:1415–1421.
  32. Sleeman MA, Watson JD, Murison JG: Neonatal murine epidermal cells express a functional multidrug-resistant pump. J Invest Dermatol 2000;115:19–23.
  33. Yano S, Ito Y, Fujimoto M, Hamazaki TS, Tamaki K, Okochi H: Characterization and localization of side population cells in mouse skin. Stem Cells 2005;23:834–841.
  34. Mills AA, Zheng B, Wang XJ, Vogel H, Roop DR, Bradley A: p63 is a p53 homologue required for limb and epidermal morphogenesis. Nature 1999;398:708–713.
  35. Yang A, Schweitzer R, Sun D, Kaghad M, Walker N, Bronson RT, Tabin C, Sharpe A, Caput D, Crum C, McKeon F p63 is essential for regenerative proliferation in limb, craniofacial and epithelial development. Nature 1999;398:714–718.
  36. Stasiak PC, Purkis PE, Leigh IM, Lane EB: Keratin 19: predicted amino acid sequence and broad tissue distribution suggest it evolved from keratinocyte keratins. J Invest Dermatol 1989;92:707–716.
  37. Lyle S, Christofidou-Solomidou M, Liu Y, Elder DE, Albelda S, Cotsarelis G: The C8/144B monoclonal antibody recognizes cytokeratin 15 and defines the location of human hair follicle stem cells. J Cell Sci 1998;111:3179–3188.
  38. Zhu AJ, Watt FM: β-Catenin signalling modulates proliferative potential of human epidermal keratinocytes independently of intercellular adhesion. Development 1999;126:2285–2298.
  39. Marchetto MC, Muotri AR, Burns DK, Friedberg EC, Menck CF: Gene transduction in skin cells: preventing cancer in xeroderma pigmentosum mice. Proc Natl Acad Sci USA 2004;101:17759–17764.
  40. Ghadially R: In search of the elusive epidermal stem cell. Ernst Schering Res Found Workshop 2005;54:45–62.
  41. Rothberg S, Crounse RG, Lee JL: Glycine-C-14-incorporation into the proteins of normal stratum corneum and the abnormal straum corneum of psoriasis. J Invest Dermatol 1961;37:497–505.
  42. Bickenbach JR, Dunnwald M: Epidermal stem cells: characteristics and use in tissue engineering and gene therapy. Adv Dermatol 2000;16:159–183; discussion 184.
  43. Fan H, Lin Q, Morrissey GR, Khavari PA: Immunization via hair follicles by topical application of naked DNA to normal skin. Nat Biotechnol 1999;17:870–872.
  44. Jensen TG: Cutaneous gene therapy. Ann Med 2007;39:108–115.
  45. Foldvari M, Kumar P, King M, Batta R, Michel D, Badea I, Wloch M: Gene delivery into human skin in vitro using biphasic lipid vesicles. Curr Drug Deliv 2006;3:89–93.
  46. Meykadeh N, Mirmohammadsadegh A, Wang Z, Basner-Tschakarjan E, Hengge UR: Topical application of plasmid DNA to mouse and human skin. J Mol Med 2005;83:897–903.
  47. Jensen TG: Gene transfer into human epidermis as an experimental model for somatic gene therapy. Dan Med Bull 2004;51:155–166.
  48. Larcher F, Dellambra E, Rico L, Bondanza S, Murillas R, Cattoglio C, Mavilio F, Jorcano JL, Zambruno G, Del Rio M: Long-term engraftment of single genetically modified human epidermal holoclones enables safety pre-assessment of cutaneous gene therapy. Mol Ther 2007;15:1670–1676.
  49. Ferrari S, Pellegrini G, Matsui T, Mavilio F, De Luca M: Gene therapy in combination with tissue engineering to treat epidermolysis bullosa. Expert Opin Biol Ther 2006;6:367–378.
  50. Mavilio F, Pellegrini G, Ferrari S, Di Nunzio F, Di Iorio E, Recchia A, Maruggi G, Ferrari G, Provasi E, Bonini C, Capurro S, Conti A, Magnoni C, Giannetti A, De Luca M: Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells. Nat Med 2006;12:1397–1402.
  51. Ortiz-Urda S, Thyagarajan B, Keene DR, Lin Q, Fang M, Calos MP, Khavari PA: Stable nonviral genetic correction of inherited human skin disease. Nat Med 2002;8:1166–1170.
  52. Costa RM, Chigancas V, Galhardo Rda S, Carvalho H, Menck CF: The eukaryotic nucleotide excision repair pathway. Biochimie 2003;85:1083–1099.
  53. Arnaudeau-Begard C, Brellier F, Chevallier-Lagente O, Hoeijmakers J, Bernerd F, Sarasin A, Magnaldo T: Genetic correction of DNA repair-deficient/cancer-prone xeroderma pigmentosum group C keratinocytes. Hum Gene Ther 2003;14:983–996.
  54. Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K, Yamanaka S: Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 2007;131:861–872.
  55. Nakagawa M, Koyanagi M, Tanabe K, Takahashi K, Ichisaka T, Aoi T, Okita K, Mochiduki Y, Takizawa N, Yamanaka S: Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts. Nat Biotechnol 2008;26:101–106.
  56. Lowry WE, Richter L, Yachechko R, Pyle AD, Tchieu J, Sridharan R, Clark AT, Plath K: Generation of human induced pluripotent stem cells from dermal fibroblasts. Proc Natl Acad Sci USA 2008;105:2883–2888.
  57. Yu J, Vodyanik MA, Smuga-Otto K, Antosiewicz-Bourget J, Frane JL, Tian S, Nie J, Jonsdottir GA, Ruotti V, Stewart R, Slukvin II, Thomson JA: Induced pluripotent stem cell lines derived from human somatic cells. Science 2007;318:1917–1920.
  58. Dupin E, Calloni G, Real C, Goncalves-Trentin A, Le Douarin NM: Neural crest progenitors and stem cells. C R Biol 2007;330:521–529.
  59. Crane JF, Trainor PA: Neural crest stem and progenitor cells. Annu Rev Cell Dev Biol 2006;22:267–286.
  60. Toma JG, Akhavan M, Fernandes KJ, Barnabé-Heider F, Sadikot A, Kaplan DR, Miller FD: Isolation of multipotent adult stem cells from the dermis of mammalian skin. Nat Cell Biol 2001;3:778–784.
  61. Toma JG, McKenzie IA, Bagli D, Miller FD: Isolation and characterization of multipotent skin-derived precursors from human skin. Stem Cells 2005;23:727–737.
  62. Sieber-Blum M, Grim M: The adult hair follicle: cradle for pluripotent neural crest stem cells. Birth Defects Res C Embryo Today 2004;72:162–172.
  63. Amoh Y, Li L, Katsuoka K, Penman S, Hoffman RM: Multipotent nestin-positive, keratin-negative hair-follicle bulge stem cells can form neurons. Proc Natl Acad Sci USA 2005;102:5530–5534.
  64. McKenzie IA, Biernaskie J, Toma JG, Midha R, Miller FD: Skin-derived precursors generate myelinating Schwann cells for the injured and dysmyelinated nervous system. J Neurosci 2006;26:6651–6660.
  65. Badiavas, EV, Falanga V: Treatment of chronic wounds with bone marrow-derived cells. Arch Dermatol 2003;139:510–516.
  66. Ferrari G, Cusella-De Angelis G, Coletta M, Paolucci E, Stornaiuolo A, Cossu G, Mavilio F: Muscle regeneration by bone marrow-derived myogenic progenitors. Science 1998;279:1528–1530.
  67. Kocher AA, Schuster MD, Szabolcs MJ, Takuma S, Burkhoff D, Wang J, Homma S, Edwards NM, Itescu S: Neovascularization of ischemic myocardium by human bone-marrow-derived angioblasts prevents cardiomyocyte apoptosis, reduces remodeling and improves cardiac function. Nat Med 2001;7:430–436.
  68. Orlic D, Kajstura J, Chimenti S, Jakoniuk I, Anderson SM, Li B, Pickel J, McKay R, Nadal-Ginard B, Bodine DM, Leri A, Anversa P: Bone marrow cells regenerate infarcted myocardium. Nature 2001;410:701–705.
  69. Metcalfe AD, Ferguson MW: Tissue engineering of replacement skin: the crossroads of biomaterials, wound healing, embryonic development, stem cells and regeneration. J R Soc Interface 2007;4:413–437.
  70. Kassem M, Kristiansen M, Abdallah BM: Mesenchymal stem cells: cell biology and potential use in therapy. Basic Clin Pharmacol Toxicol 2004;95:209–214.
  71. Jiang Y, Jahagirdar BN, Reinhardt RL, Schwartz RE, Keene CD, Ortiz-Gonzalez XR, Reyes M, Lenvik T, Lund T, Blackstad M, Du J, Aldrich S, Lisberg A, Low WC, Largaespada DA, Verfaillie CM: Pluripotency of mesenchymal stem cells derived from adult marrow. Nature 2002;418:41–49.
  72. Abe R, Donnelly SC, Peng T, Bucala R, Metz CN: Peripheral blood fibrocytes: differentiation pathway and migration to wound sites. J Immunol 2001;166:7556–7562.
  73. Yang L, Scott PG, Giuffre J, Shankowsky HA, Ghahary A, Tredget EE: Peripheral blood fibrocytes from burn patients: identification and quantification of fibrocytes in adherent cells cultured from peripheral blood mononuclear cells. Lab Invest 2002;82:1183–1192.
  74. Bellini A, Mattoli S: The role of the fibrocyte, a bone marrow-derived mesenchymal progenitor, in reactive and reparative fibroses. Lab Invest 2007;87:858–870.

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