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Human embryonic stem cells derivatives enable full reconstruction of the pluristratified epidermis.
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Edité par CCSD -
Background: Cell therapy for large burns has successfully made use of autologous epidermis reconstructed in vitro for more than two decades. One limit to the efficacy of the current procedures is the need for a long lasting expansion of the patient's own keratinocytes. Immediate access to unlimited amounts of human epidermis may contribute to protect patients awaiting autologous grafts. The keratinocyte progeny of human embryonic stem cells is an interesting candidate for such a role, provided they are fully capable of reconstructing a pluristratified epidermis. Methods: Human embryonic stem cells were seeded on fibroblasts feeder cells during forty days in a medium supplemented with BMP4 (0.5 nM) and ascorbic acid (0.3 mM). A molecular characterization of cell differentiation was performed all along 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. Finding: Using a protocol that respected the chronobiology of epidermis formation during human ontogenesis, we have generated a homogenous population of cells exhibiting all phenotypic characteristics of basal keratinocytes from hESC. These cells readily constructed a pluristratified epidermis displaying normal human characteristics after seeding on an artificial matrix, and either secondarily grown in vitro or else transplanted in immunodeficient mice. Interpretation: Human embryonic stem cells can be coaxed to differentiate into basal keratinocytes that are fully functional, i.e. construct a pluristratified epidermis. This resource may be developed to provide temporary skin substitutes to patients awaiting autologous grafts.
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