The objective of the Tissue Culture Core is to provide a common resource which will provide specifically characterized cell cultures. By centralizing this effort to one facility, this should expedite our efforts to carry out the specific aims of each component project in a timely manner. As we have successfully done in the past, each cell type cultured in the facility will be screened and well defined. This facility will also facilitate interactions between the various projects. The overall contribution of this kind of service is apparent. Firstly, the economic advantage via centralization of commonly used reagents, vascular tissue, and manpower hours is considerable. This clearly results in reduction of supplies and salaries which would be expended in multiple laboratories to achieve similar objectives. Secondly, the provision of common experimental regimens and cells to all investigators in the program allows for consistency between different laboratories. The Tissue Culture Core facility will provide a wide spectrum of cell types and materials in a defined manner on a routine basis, including human arterial smooth muscle cells, adventitial fibroblasts, umbilical vein endothelial cells, rabbit and murine arterial smooth muscle cells, saphenous vein smooth muscle cells, human monocyte-like cell lines MM6, RAW 264.7, THP-1 and human peripheral blood monocytes. Fresh human platelets and neutrophils will also be isolated as needed by individual investigators. All cultures are characterized immunocytochemically and routinely monitored for mycoplasma contamination. The Core also will provide media and sera to the individual investigators when needed. The Core will be a resource for the growth and metabolic characterization of specific cell types, and the Core provides advice, expertise, and defined protocols as needed. The Core will perform LDH (lactate dehydrogenase) activity assays on cells as a measure of cell viability in response to the experimental conditions outlined by the individual component projects. Trypan blue exclusion evaluations will also be carried out as a rapid screening method. Currently, we have a well-functioning Tissue Culture facility, containing two five-foot Sterilgard laminar flow hoods, cell storage receptacle, an inverted, Nikon microscope, a confocal microscope, and three double Forma- Scientific water-jacketed flowing CO/2 incubators. There are also two centrifuges in this facility.
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