The Cell Culture Core is to serve several main functions. It will be a cost-effective, time-saving resource for a) freshly cultured and immortalized skin cells and skin equivalents; b) genetically modified skin cells; c) specialized media; d) antibodies and molecular probes for detection of skin specific markers. In addition, it will be a center for: a) information and advice in cell culture techniques; and b) development of new techniques applicable to skin research, such as optimizing procedures for efficient gene delivery into fastidious cells (endothelial cells and melanocytes) and tissue engineering that can be used for wound healing. The latter function will be performed in collaboration with the SCID mouse/Human Xenograft Core and the Tissue Aquisition & Analysis Core of the YSDRCC. The core will provide: 1. Primary cultures of normal human skin ceils (melanocytes, keratinocytes, fibroblasts and endothelial cells) from healthy donors, as well as malignant, mutant and other abnormal cells from the same patient or different patients; 2. Primary and immortalized mouse cells; 3. Genetically modified mouse melanocytes keratinocytes and fibroblasts; 4. Co-cultures, e.g., keratinocytes/melanocytes, fibroblasts/melanocytes, fibroblasts/keratinocytes; endothelial cells/keratinocytes; and various combinations thereof; 5. Skin equivalents; 6. Large volumes of cells for specified procedures such as the construction of cDNA libraries, analysis of genetic alterations, global gene expression by microarrays (mRNA), protein purification, proteomics, identification of cell surface proteins; 7. Advice and hands-on training for the growing of cells especially those that have so far proven fastidious, such as CTCL and melanocytes from primary melanomas from the radial growth phase. Significance: 1. The core provides cells to individual investigators who lack the expertise or equipment, and to others who wish to save the labor-intensive process. Both groups enjoy the highly economical and cost effectiveness of our services; 2. This core improves the consistency, ease and reproducibility of obtaining skin cells and skin equivalents, and thus enhances the quality of research and reduces the probability of artifacts due to the presence of mycoplasma and/or contaminating cells; 3. The expertise in the central facility provides advice and hands-on help to scientists in growing normal cells that otherwise have been known to be recalcitrant; 4. The core brings together scientists with common interests and enhances interactions that otherwise would not materialize; 5. The core will enhance the formation of new core facilities, such as imaging; 6. The core provides primary cultures and special cell types to investigators outside of Yale.
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