Embryonic Stem Cells and Squamous Epithelia

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DESCRIPTION (provided by applicant): Human embryonic stem (ES) cells are now available for the study of the generation of somatic cell types. For this purpose, human ES cells have important advantages over the murine ES cells available earlier. We propose to study the generation of keratinocytes of stratified squamous epithelium from the human ES cells (WA01). ES cells are known to have the capacity to generate such epithelium outside the developing embryo, since the epithelium develops when the ES cells are injected into scid mice. But our main intention is to find conditions promoting differentiation of the ES cells in culture and then to isolate strains of keratinocytes resulting from that differentiation, using 3T3 support. We will examine the factors that influence the development of keratinocytes, using as criterion a quantitative measure of the number obtained, and in this way arrive at the conditions producing maximal yield of keratinocytes. We will identify, when possible, the specific squamous epithelium to which isolated keratinocytes belong. We will attempt to find somatic stem cell precursors of the mature keratinocytes using stem cell markers for their identification. Depending on the nature of the keratinocytes or keratinocyte precursors isolated, practical applications can be readily envisioned.