CORE--TISSUE CULTURE

组织培养核心的目标是提供一个共同的资源，该资源将提供具有特定特征的细胞培养。通过将这一努力集中到一个设施，这将加快我们及时实现每个组成部分项目的具体目标的努力。在过去的10年里，这种类型的Core对我们来说运行得非常好，我们希望继续这样做。正如我们在过去成功做到的那样，在该设施中培养的每一种细胞类型都将经过筛选和充分鉴定。这一设施将提供各种项目之间的互动。这类发球的总体贡献是显而易见的。首先，通过将常用试剂、血管组织和人力时间集中起来，经济优势是相当大的。这显然导致用品和工资的减少，这将花费在多个实验室，以实现类似的目标。其次，向项目中的所有研究人员提供共同的实验方案和细胞，允许不同实验室之间的一致性。组织培养核心设施将以常规方式提供广泛的细胞类型和材料，包括人动脉平滑肌细胞、外膜成纤维细胞、脐静脉内皮细胞；兔和小鼠动脉平滑肌细胞；人单核细胞样细胞系MM6、RAW 264.7、THP-1和人外周血单核细胞。新鲜的人体血小板和中性粒细胞将根据个别研究人员的需要进行分离。目前所有培养物的特征都是免疫细胞化学，并常规监测支原体污染。中心还将在需要时向个别调查人员提供媒体和血清。核心将是特定细胞类型的代谢特性的生长资源，核心将根据需要提供建议、专业知识和定义的细胞培养方案。核心将对细胞进行乳酸脱氢酶(LDH)活性分析，以此作为细胞活力的衡量标准，以响应各个组件项目概述的实验条件。台盼蓝排除评估也将作为一种快速筛查方法进行。目前，我们拥有运转良好的组织培养设施，包括三个五英尺长的Sterilgard层流罩、四个细胞存储容器、一个倒置尼康显微镜、一个共焦显微镜和三个双层Forma-Science水套流动二氧化碳孵化器。这个设施里还有两台离心机。

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.