Generation of keratinocytes from normal and recessive dystrophic epidermolysis bullosa-induced pluripotent stem cells

Abstract

Embryonic stem cells (ESCs) have an unlimited proliferative capacity and extensive differentiation capability. They are an alternative source for regenerative therapies with a potential role in the treatment of several human diseases. The clinical use of ESCs, however, has significant ethical and biological obstacles related to their derivation from embryos and potential for immunological rejection, respectively. These disadvantages can be circumvented by the alternative use of induced pluripotent stem cells (iPSCs), which are generated from an individual's (autologous) somatic cells by exogenous expression of defined transcription factors and have biological characteristics similar to ESCs. In recent years, patientspecific iPSCs have been generated to study disease mechanisms and develop iPSC-based therapies. The development of iPSC-based therapies for skin diseases requires successful differentiation of iPSCs into cellular components of the skin, including epidermal keratinocytes. Here, we succeeded in generating iPSCs not only from normal human fibroblasts but also from fibroblasts isolated from the skin of two patients with recessive dystrophic epidermolysis bullosa. Moreover, we differentiated both of these iPSCs into keratinocytes with high efficiency, and generated 3D skin equivalents using iPSC-derived keratinocytes, suggesting that they were fully functional. Our studies indicate that autologous iPSCs have the potential to provide a source of cells for regenerative therapies for specific skin diseases.reprogramming | retinoic acid | bone morphogenetic protein 4 | cellular therapy

Impaired epithelial differentiation of induced pluripotent stem cells from ectodermal dysplasia-related patients is rescued by the small compound APR-246/PRIMA-1 MET

“…(Magnification: B, 40×; D, 20×.) BMP-4-based protocols have been established to obtain keratinocytes from pluripotent cells (12,14,25) and very recently, Itoh et al used BMP-4 combined with retinoic acid for a limited time to generate keratinocytes from iPSC from dystrophic epidermolysis bullosa patients reaching 30-40% of K14 + cells that could be enriched only after passaging or cell sorting (26). Despite possible enrichment and satisfactory 3D potential (14,26), improved protocols for highly homogenous production of keratinocytes from pluripotent cells are still needed to consider cell therapy.…”

Site-specific genome editing for correction of induced pluripotent stem cells derived from dominant dystrophic epidermolysis bullosa

“…S4). iPSC-derived keratinocytes and fibroblasts were generated as described previously (20,21). No differences were detected among normal, untreated DDEB, and gene-edited DDEB iPSC lines in the process of differentiation into keratinocytes and fibroblasts.…”

“…iPSC-derived keratinocytes and fibroblasts were generated as described previously (20,21). These iPSCderived keratinocytes and fibroblasts were characterized by immunofluorescence studies as described previously (Table S3) (20,21).…”

In vitro skin three-dimensional models and their applications

“…Disease models can be generated with iPSCs derived from somatic cells having a genetic mutation. To mimic keratinocytes in recessive dystrophic epidermolysis bullosa, Itoh et al isolated cells from diseased patients, reprogrammed them into iPSCs and differentiated them into keratinocytes [94]. Prospectively, iPSCs have the potential to predict patient-specific response to therapies.…”