CORE--TISSUE CULTURE

点击翻译按钮获取中文摘要

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 will expedite our efforts to carry out the specific aims of each component project in a timely manner. This type of Core has worked very well for us in the past 10 years, and we would like to continue it. As we have successfully done in the past, each cell type cultured in the facility will be screened and well- characterized. This facility will provide interactions between the various projects. The overall contribution of this type of serve is apparent. Firstly, the economic advantage via centralization of commonly used reagents, vascular tissue, and man-power 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 protocols 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 of routine basis, including human arterial smooth muscle cells, adventitial fibroblasts, umbilical vein endothelial cells; rabbit and murine arterial 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 be isolated as needed by individual investigators. All cultures are currently 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 in metabolic characterization of specific cell types, and the Core will provide advice, expertise, and defined cell culture 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 three five-foot Sterilgard laminar flow hoods, four cell storage receptacles, an inverted Nikon microscope, a confocal microscope, and three double Forma-Scientific water-jacketed flowing CO2 incubators. There are also two centrifuges in this facility.