Differentiation of Mouse Induced Pluripotent Stem Cells into a Multipotent Keratinocyte Lineage

Abstract

Recent breakthroughs in the generation of induced pluripotent stem cells (iPSCs) have provided a novel renewable source of cells with embryonic stem cell-like properties, which may potentially be used for gene therapy and tissue engineering. Although iPSCs have been differentiated into various cell types, iPSC-derived keratinocytes have not yet been obtained. In this study, we report the in vitro differentiation of mouse iPSCs into a keratinocyte lineage through sequential applications of retinoic acid and bone-morphogenetic protein-4 and growth on collagen IV-coated plates. We show that iPSCs can be differentiated into functional keratinocytes capable of regenerating a fully differentiated epidermis, hair follicles, and sebaceous glands in an in vivo environment. Keratinocytes derived from iPSCs displayed characteristics similar to those of primary keratinocytes with respect to gene and protein expression, as well as their ability to differentiate in vitro and to reconstitute normal skin and its appendages in an in vivo assay. At present, no effective therapeutic treatments are available for many genetic skin diseases. The development of methods for the efficient differentiation of iPSCs into a keratinocyte lineage will enable us to determine whether genetically corrected autologous iPSCs can be used to generate a permanent corrective therapy for these diseases.

Natural Gene Therapy in Dystrophic Epidermolysis Bullosa

“…22 Keratinocytes and fibroblasts can be reprogrammed into induced pluripotent stem cells (iPSCs) and subsequently differentiated into keratinocytes, providing an unlimited source of autologous keratinocytes. 23 Recently, 2 groups described the generation of iPSCs from the cells of patients with RDEB and the subsequent spontaneous 24 and directed 25 differentiation of these iPSCs into keratinocytes. The use of revertant cells obviates the need for potentially dangerous virus-mediated gene correction 26 and circumvents the risk of immunologic rejection, and, if combined with the patient-specific iPSCs approach, provides the opportunity to grow a large number of healthy skin grafts.…”

Epidermal Differentiation in Barrier Maintenance and Wound Healing

“…In fact, procedures have been developed to effectively differentiate iPSCs into various cell types, including functional keratinocytes capable of reconstituting a normal stratified epidermis, hair follicles, and sebaceous glands. 63 Mesenchymal stem cells. In addition to resident SCs in the skin, mesenchymal stem cells (MSCs) derived from either the bone marrow or adipose tissue have been shown to accelerate cutaneous wound regeneration.…”

Novel and emerging therapies in the treatment of recessive dystrophic epidermolysis bullosa

“…It is possible to correct RDEB fibroblasts through homologous recombination using transcription activator-like effector nucleases (TALENs) and then reprogram these into iPSCs, which then differentiate into keratinocytes (135). Murine studies have also successfully generated iPSCs in culture from multipotent keratinocyte lineages capable of forming a fully developed epidermis (136). Subsequently, others have reported successful generation of iPSCs from healthy human skin fibroblasts and individuals with RDEB (137).…”