A Cell Culture Core is proposed to serve as a central facility for both quality controlled material resources and expert technical support in cell culture to investigators of the UWSDRC. This core will facilitate efficient execution of projects involving the use and study of skin cells from normal individuals and patients with skin diseases and murine genetic models of human skin disorders. The specific functions of this core are: 1) Provide quality-assured cultures of the three major cell types of the skin- keratinocytes, fibroblasts and melanocytes from normal and diseased human skin and mouse models of human skin disorders. These are important and critical resources for cell and molecular studies on cell death and differentiation. The Core will culture, expand and cryopreserve the 3 cell types as matched sets, when available, that will serve as isogenic controls. 2) Generate induced pluripotent stem cells (iPSCs) from skin cells. Skin derived iPSCs are useful for studies involving regulation of skin cell differentiation and modeling genetic skin diseases in vitro. The differentiated cells can be studied either as monolayers or after their incorporation into 3-D organotypic cultures. The Core will provide the required material resources and technical expertise to investigators and hands-on training, as needed. 3) Generate quality controlled lentiviruses. High titer lentiviruses have become indispensable reagents for effective gene transfer into primary cells. The UWSDRC Cell Culture Core will provide both material resources such as high quality packaging cells and technical expertise for production of viruses. 4) Provide technical expertise and hands-on training to investigators in manipulation of skin cells in culture. Many studies of cell death and differentiation involve transfection/electroporation with plasmid DNA or small inhibitory RNA or transduction with lentiviruses. 5) Provide cultured cells as source for macromolecules. Biochemical and molecular biological studies of cell death and differentiation of skin cells often require macromolecules (DNA, RNA or protein) isolated from individual cell types. The Core will provide frozen or fresh cell pellets for extraction of macromolecules of interest.
Skin diseases constitute a major public health burden in the US and worldwide. Understanding the cellular and molecular basis of these diseases is facilitated by availability of quality controlled pure populations of skin cells derived from both healthy and affected individuals. The Cell Culture Core proposed here will serve a large interdisciplinary group of scientists in their studies of human skin biology in health and disease.
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