Human embryonic stem-cell derivatives for full reconstruction of the pluristratified epidermis: a preclinical study

The Lancet, 374(9703):1745-1753

GUENOU H., NISSAN X., LARCHER F., FETEIRA J., LEMAITRE G., SAIDANI M., DEL RIO M., BARRAULT C., BERNARD FX., PESCHANSKI M., BALDESCHI C. and WAKSMAN G. (2009)

INSERM/UEVE U-861, I-STEM, AFM, Institute for Stem Cell Therapy and Exploration of Monogenic diseases, Evry Cedex, France.
CIEMAT–CIBER-ER, Epithelial Biomedicine Division, Madrid, Spain.
Bioalternatives, Gençay, France.

Abstract

Cell therapy for large burns is dependent upon autologous epidermis reconstructed in vitro. However, the effectiveness of current procedures is limited by the delay needed to culture the patient’s own keratinocytes. To assess whether the keratinocyte progeny of human embryonic stem-cell (hESCs) could be used to form a temporary skin substitute for use in patients awaiting autologous grafts, we investigated the cells’ capability of constructing a pluristratified epidermis.
hESCs from lines H9 and SA01 were seeded at least in triplicate on fibroblast feeder cells for 40 days in a medium supplemented with bone morphogenetic protein 4 and ascorbic acid. Molecular characterisation of cell differentiation was done throughout the process by quantitative PCR, fluorescence-activated cell sorting, and immunocytochemical techniques. Keratinocyte molecular differentiation and functional capacity to construct a human epidermis were assessed in vitro and in vivo.
From hESCs, we generated a homogeneous population of cells that showed phenotypic characteristics of basal keratinocytes. Expression levels of genes encoding keratin 14, keratin 5, integrin alpha6, integrin beta4, collagen VII, and laminin 5 in these cells were similar to those in basal keratinocytes. After seeding on an artificial matrix, keratinocytes derived from hESCs (K-hESCs) formed a pluristratified epidermis. Keratin-14 immunostaining was seen in the basal compartment, with keratin 10 present in layers overlying the basal layer. Involucrin and filaggrin, late markers of epidermal differentiation, were detected in the uppermost layers only. 12 weeks after grafting onto five immunodeficient mice, epidermis derived from K-hESCs had a structure consistent with that of mature human skin. Human involucrin was appropriately located in spinous and granular layers and few Ki67-positive cells were detected in the basal layer.
hESCs can be differentiated into basal keratinocytes that are fully functional–ie, able to construct a pluristratified epidermis. This resource could be developed to provide temporary skin substitutes for patients awaiting autologous grafts.

© 2009 Elsevier Ltd.

KEYWORDS: Human embryonic stem-cell ; hESCs; Keratinocyte

Check out Bioalternatives’ updates and experience new testing ideas

  • Bioassays, models and services
  • Posts and publications
  • Events

Related Posts or publications

Acné vulgaris : Origines, symptômes et traitements actuels L’acné vulgaris est une maladie inflammatoire chronique cutanée qui touche la plupart des adolescents et concerne plus de la moitié des consultations en dermatologie. Cette pathologie, caractérisée par une hypersécrétion de sébum peut être à l’origin...
Acné vulgaris : Physiopathologie et mécanismes cellulaires L’acné vulgaris est une pathologie inflammatoire chronique cutanée localisée au niveau du follicule pilo-sébacé. Elle apparaît suite à une surproduction de sébum liée à des dysfonctionnements d’origine hormonale (androgène), environnementale (polluti...
La glande sébacée : Physiologie et fonctions La glande sébacée est un organe localisé au niveau du derme. Elle est responsable de la synthèse et de la sécrétion du sébum qui entre dans la composition du film hydrolipidique. Le rôle du sébum est de protéger la peau des agressions extérieures et ...
Interleukin-17A-induced production of acute serum amyloid A by keratinocytes contributes to psoriasis pathogenesis Acute-serum Amyloid A (A-SAA), one of the major acute-phase proteins, is mainly produced in the liver but extra-hepatic synthesis involving the skin has been reported. Its expression is regulated by the transcription factors NF-κB, C/EBPβ, STAT3 acti...
Development of a new model of reconstituted mouse epidermis and characterization of its response to proinflammatory cytokines The development of three-dimensional models of reconstituted mouse epidermis (RME) has been hampered by the difficulty to maintain murine primary keratinocyte cultures and to achieve a complete epidermal stratification. In this study, a new protocol ...