| Summary: | Studies on mouse keratinocyte stem cells are hampered by the absence of markers for epidermal stem cells, and there is little information about the <I>in vivo</I> or <I>in vitro</I> micro environment required for maintenance of the murine keratinocyte stem cell. In this study, a unique immortal mouse follicular dermal papilla cell line was generated in order to investigate the interactions <I>in vitro</I> and <I>in vivo</I> between dermal papilla cells and keratinocytes. The cell line was characterised and used for experimental analysis of keratinocyte self-renewal. Characterisation of the DPC1" line included growth properties, morphology and expression at mRNA level of growth factors known to be important in keratinocyte or dermal papilla cell growth. Primary mouse epidermal keratinocytes were cultured in monolayer on the dermal papilla cell line (DPC1") and compared to cultures on other fibroblast lines (A31 3T3 and J2 3T3). The DPC1" supported a more proliferative phenotype of keratinocyte, both in primary culture and after subculture. The keratinocytes were also grown in organotypic culture and the DPC1" cells supported the formation of a multilayered proliferative epidermis-like structure. <I>In vivo</I> it was shown that the keratinocytes were able to reform a self-renewing epidermis in the graft bed when they were grafted in conjunction with DPC1". Finally, keratinocytes were cultured on the protein matrix synthesised by the DPC1" and it was demonstrated that there was greater attachment and proliferation on this matrix when compared to matrix derived from other cell lines. It is proposed that the DPC1" cell line can reproduce at least part of the micro environments required in terms of the protein matrix and growth factors for maintenance of the mouse keratinocyte stem cell.
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