The function of the cell culture core will be to provide strictly defined cells in culture for the participating investigators. The cell culture core will be responsible for the the isolation of primary cultures of human dermal microvascular endothelial cells, murine blood outgrowth endothelial cells (BOEC), murine smooth muscle cells, murine pulmonary vein and artery endothelial cells and murine fibroblasts. Cell lines including murine liver cells and human pulmonary vein and artery endothelial cells, will also be cultured. Primary cells will be grown in endotoxin-free conditions utilizing experienced cell culture techniques. The.purity of the cells will be verified using a variety of immunological techniques and functional assays. The cell culture core will distribute plates of cells to all investigators for the studies outlined in their proposals.The laboratory has rigorous techniques to provide endotoxin-free cell cultures to minimize cell activation, a critical criteria for many studies. This is accomplished by using endotoxin-free media and regular testing of all reagents used for cell culture for endotoxin using the limulus amoebocyte lysate assay. The P.I. and technician have many years of experience growing endothelial and other cells in culture. Sickle cell anemia is a devastating disease manifest by anemia, painful crises and organ injury such as stroke. The cells cultured in this Core will be used by investigators to test new treatments. By using cells, safety and effectiveness can be studied before giving new therapies to humans.
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