Analysis of transferred keratinocyte-like cells derived from mouse embryonic stem cells on experimental surgical skin wounds of mouse
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Date
2013
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Abstract
Autologous/allogenic skin grafts constituted from differentiated adult or embryonic stem cells can be used in treatment of skin disorders. In our study we aimed to differentiate keratinocytes from mouse embryonic stem cells and the transfer of viable keratinocyte-like cells to a model of surgical skin wound of mouse. Embryoid bodies, derived from mouse embryonic stem cells, were cultured on basement membrane matrix with added BMP-4 for 10 days. The identification of differentiated keratinocyte-like cells was done by detection of cytokeratin-8 and cytokeratin-14 localization using an indirect immunoperoxidase technique and transmission electron microscopy evaluation. Distribution of BrdU, cytokeratin-8 and cytokeratin-14 were evaluated using an indirect immunoperoxidase technique from the experimental (dressing including BrdU labelled cells applied after the surgical wound was created on mouse), control (only the surgical wound was created on mouse) and sham (only the dressing applied after the surgical wound was created on mouse) in groups after 3, 5 and 7 days. Immunohistochemically and ultrastructurally, cells derived from mouse embryonic stem cells were similar to differentiated keratinocyte-like cells. Differentiated keratinocyte-like cells were demonstrated by positive BrdU, cytokeratin-8 and cytokeratin-14 staining after transfer to the wound area. In the experimental group wound healing was better after transferring differentiated keratinocytes when compared to the sham and control groups. In vivo continuity and usability of derived cells are very important issues. In wound repair mechanisms, keratinocyte-like cells could provide positive effects during the wound healing and could be used in clinical treatments of wound repair process. © 2012 Elsevier GmbH.
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Keywords
Animals , Cell Differentiation , Dermatologic Surgical Procedures , Embryonic Stem Cells , Keratinocytes , Male , Mice , Mice, Inbred BALB C , Wound Healing , bone morphogenetic protein 4 , cytokeratin 14 , cytokeratin 8 , peroxidase , animal cell , animal experiment , animal model , animal tissue , article , basement membrane , biotinylation , cell differentiation , cell function , cell proliferation , cell transfer , cell viability , controlled study , embryo , embryoid body , embryonic stem cell , immunohistochemistry , immunolocalization , immunoreactivity , keratinocyte , male , microscopy , mouse , neutrophil , nonhuman , skin injury , transmission electron microscopy , wound healing