In contrast to most mammalian organs, many cellular elements of normal human skin can now be cultured. Techniques to culture human keratinocytes (first evolved by HSDRC member Dr. Green), dermal fibroblasts, melanocytes, and microvascular endothelial cells are now standardized, and most of these cells can be obtained from at least one commercial vendor (e.g., Clonetics). Each cell type requires different medium, however, and it is impractical for one laboratory to grow all such cells. Furthermore, unless study of one of these individual cell in a consistent and detailed fashion, it is impractical to initiate the culture of such cells on a regular basis. The main goal of the Cell Culture core is to facilitate experimental use of freshly cultured skin cells, of both human and murine origin. With regards to murine cells, particular emphasis will be placed on the culturing of transgenic-derived keratinocytes, melanocytes, dermal fibroblasts and endothelial cells.
The specific aims of the proposal are to provide the following services: 1) Procurement of human and murine skin. 2) Special media preparation for specific cell types. 3) Isolation and cultivation of skin-derived primary cells, including large scale preparations for biochemical studies (such as RNA and cDNA library preparation; cytokine and enzyme purifications, etc.). 4) Training in the in vitro isolation of cellular elements of skin, such that investigators appropriately trained and having access to appropriate media can initiate and maintain cell cultures in their laboratories.
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