Flow Cytometry and Cell Sorting Shared Resource Facility

项目总结/摘要 流式细胞术和细胞分选共享资源设施（FCCS SRF）促进高影响癌症 Markey癌症中心（MCC）内的科学，并得到目前CCSG资金的资助。FCCS SRF 自1983年以来一直在肯塔基州（英国）大学持续运作，并在唐纳德的指导下 科恩博士，1992年至今。虽然是英国的一个机构核心设施，但在CCSG的当前期间， 68%的用户是MCC成员，53%的用户是经过同行评审的MCC成员。到 为了支持MCC成员的研究工作，FCCS SRF提供最先进的流式细胞术服务， 全面的免疫监测服务，支持所有4个研究项目的癌症研究。各种 流式细胞仪服务，包括多参数细胞表型分析（最多同时使用12种生物标志物）， 允许用户分析表面和细胞内生物标志物、DNA含量、细胞周期、凋亡和坏死， 细胞因子/生长因子合成、信号转导和细胞活化。在管理协委会领导的指导下， FCCS SRF最近扩大了服务范围，为MCC成员提供全面的免疫监测， 人类基础和临床癌症研究。免疫监测服务包括处理新鲜血液， 用于分离免疫和非免疫细胞以进行功能测定和冷冻保存的肿瘤组织， 将细胞建库以供以后分析。银行服务是与生物样本合作提供的。 采购和转化病理学SRF和癌症研究信息学SRF。新鲜分离 并且可以分析储存的细胞样品的许多细胞功能， 细胞因子/生长因子/其他可溶性因子，通过ELISA、ELISPOT和Luminex多路复用，除了流动 细胞计数方法。还提供了定量抗原特异性T细胞增殖、细胞毒性 抗体合成和细胞因子的产生。蜂窝产品服务包括生成 来自血液和/或肿瘤组织的树突细胞和巨噬细胞以及来自血液和/或肿瘤组织的T细胞扩增 肿瘤组织FCCS与生物统计学和生物信息学SRF合作，提供高维数据 用于鉴定复杂细胞的多参数流式细胞术和Luminex多分析物谱分析 群体和稀有细胞亚群。在上一个CCSG资助期间，FCCS服务有助于 在过去四年中资助了118份出版物和49笔赠款，并启动了协商与合作 MCC成员的4项临床试验。全面的流式细胞仪和免疫监测服务 由FCCS提供，结合FCCS免疫测定设计服务，为MCC研究人员提供 有机会询问新的癌症生物标志物和免疫系统的相互作用， 参与MCC的基础和临床癌症研究。

PROJECT SUMMARY/ABSTRACT The Flow Cytometry and Cell Sorting Shared Resource Facility (FCCS SRF) facilitates high impact cancer science within the Markey Cancer Center (MCC) and is aided by current CCSG funding. The FCCS SRF has been in continuous operation at the University of Kentucky (UK) since 1983 and under the direction of Donald Cohen, PhD, since 1992. Although an institutional core facility at UK, during the current period of CCSG funding, 68% of users were MCC members, and 53% of users were peer-reviewed funded MCC members. To support research efforts of MCC members, the FCCS SRF provides state-of-art flow cytometric services and comprehensive immune monitoring services to support cancer research by all 4 Research Programs. A variety of flow cytometric services, including multi-parametric cell phenotyping (up to 12 biomarkers simultaneously), allows users to analyze surface and intracellular biomarkers, DNA content, cell cycle, apoptosis and necrosis, cytokine/growth factor synthesis, signal transduction and cell activation. Under guidance by MCC leadership, the FCCS SRF recently expanded services to provide MCC members comprehensive immune monitoring for human basic and clinical cancer research. Immune monitoring services include processing fresh blood and tumor tissue for isolation of immune and non-immune cells for functional assay and cryopreservation and banking of cells for later analysis. Banking services are provided in collaboration with the Biospecimen Procurement and Translational Pathology SRF and the Cancer Research Informatics SRF. Freshly isolated and banked cell samples can be analyzed for a number of cellular functions including synthesis of cytokines/growth factors/other soluble factors by ELISA, ELISPOT and Luminex multiplexing, in addition to flow cytometric approaches. Assays are also offered to quantify antigen-specific T cell proliferation, cytotoxicity, antibody synthesis and cytokine production in human systems. Cellular product services include generation of dendritic cells and macrophages from blood and/or tumor tissue and expansion of T cells from blood and/or tumor tissue. In collaboration with the Biostatistics and Bioinformatics SRF, FCCS offers high dimensional data analysis for multi-parameter flow cytometry and Luminex multi-analyte profiling for identification of complex cell populations and rare cell subsets. Over the previous CCSG funding period, FCCS services have contributed to 118 publications, 49 grants funded during the last four years, and has initiated consultation and collaboration on 4 clinical trials by MCC members. The comprehensive flow cytometry and immune monitoring services offered by FCCS, in combination with FCCS immune assay design services, are providing MCC researchers with the exceptional opportunities to interrogate novel cancer biomarkers and the immune system interactions in studies involving basic and clinical cancer research at the MCC.