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

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 ...
in vitro modeling of skin photoaging: development of evaluation tools for cosmetics Development of in vitro models skin for better understand the modifications during photo-aging induced by repetitive UV exposure.
Development of a new model of reconstructed aged skin useful to study antiageing effects of cosmetic compounds The development of new anti-ageing products needs performant in vitro models mimicking morphological changes and physiological modifications appearing during skin ageing. In order to have access to a simple model mimicking the epidermis ageing but in...
Wound healing, granulation phase and maturation phase The third phase of wound healing, consisting in the replacement of the provisional  fibrin matrix with granulation tissue once the wound has been debrided, includes several sub-phases: re-epithelialization, fibroplasia, collagen deposition and angio...