COREB: TISSUE CULTURE CORE LABORATORY The objective of the tissue culture laboratory is to support the investigators of the Program Project who use cell models and, as such, it is an integral part of all the projects in the current program. In addition to providing expertise and training, the tissue culture laboratory provides essential equipment and dedicated space that is ideally suited and maintained for successful preparation of cell cultures. Such dedicated space and equipment is not otherwise available to investigators in Projects 1 and 2;moreover, only the tissue culture laboratory is equipped to provide large numbers of cells for experiments proposed in project 3 for in vivo assay of reverse cholesterol transport. The laboratory provides numerous services e.g. standardized protocols, materials and reagents for cell culture;a ready supply of widely used cell lines;routine preparation of primary cells such as mouse peritoneal macrophages, personnel training;development of new methods; procurement and storage of cell lines;transient and stable transfection of cells;and Western blots. Thus, a centralized Tissue Culture Core facility has several advantages: 1) it conserves space, eliminates the need for redundant equipment and allows for the cost effective and timely acquisition of supplies;2) it ensures standardization of procedures and cell lines;3) it facilitates the introduction of new cell models into the program;4) serves as a training facility. Core B usage in the present grant cycle has been extensive. Over the past year the core has, on a weekly average, provided cells, reagents and materials for the following cell- based experiments: 3 efflux experiments, 1RCT experiment, and 2 primary mouse macrophage experiments. The laboratory has also provided weekly Western blots for detection of SR-BI, ABCA1 and more recently ABCG1. Core B has also recovered cells from frozen stocks for use by investigators and provided cells for studies of HDL biogenesis at least 2 times per month. We expect this pattern of use to continue in the current cycle and anticipate the following core usage distribution: Project 1- 55%;Project 2-15% and Project 3-30 %.
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