Using embryonic stem cells to successfully induce intact epidermis
According to a research report published in the journal Lancet on November 21, the I-STEM * Institute (I-STEM / Inserm UEVE U861 / AFM) team led by Marc Peschanski successfully used human embryonic stem cells to generate a complete epidermis . The research group hopes that this stem cell can be used as an alternative therapy for patients with third degree burns and patients with hereditary skin diseases in the future.
Human embryonic stem cells (hES) have two basic characteristics: the ability to proliferate indefinitely and the pluripotency to differentiate into various cells of the body.
First, the research team obtained skin stem cells-keratinocytes (keratinocytes), keratinocytes can continuously update the skin; then the researchers tested the isolated keratinocytes in vitro and organisms to form functional epidermal cells Ability.
With the support of cell biology and pharmacology methods, it is possible to transform hES cells into epidermal cells. The researchers successfully used keratinocytes to reconstruct an epidermis (epidermis) in vitro. Finally, the researchers also need to formally transfer the results obtained in vitro to the organism. In the final stage of the experiment, the researchers transplanted the obtained epidermis to mice with reduced immunity, and 12 weeks after the transplantation, the local skin area of ​​the transplanted epidermis of the mice completely returned to normal. (Bioon.com)
Bio Valley recommends the original source:
The Lancet, Volume 374, Issue 9703, Pages 1745-1753, 21 November 2009
Human embryonic stem-cell derivatives for full reconstruction of the pluristratified epidermis: a preclinical study
Hind Guenou PhD a, Xavier Nissan a, Fernando Larcher PhD b, Jessica Feteira a, Gilles Lemaitre PhD a, Manoubia Saidani a, Marcela Del Rio PhD b, Christine C Barrault c, Fran? Ois-Xavier Bernard PhD c, Marc Peschanski MD a, Dr Christine Baldeschi PhD a, Prof Gilles Waksman PhD a
Background
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 patients own keratinocytes. To assess whether the keratinocyte progeny of human embryonic stem cells (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.
Methods
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.
Findings
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 α6, integrin β4, 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.
Interpretation
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.
Funding
Institut National de la Santé et de la Recherche Médicale, University Evry Val dEssonne, Association Fran? Aise contre les Myopathies, Fondation René Touraine, and Genopole.
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