ADENOVIRUS/CARDIOMYOCYTES

核心将提供必要的支持，创作，选择，人物塑造，制作和 重组腺病毒的纯化及新生大鼠心肌细胞的分离培养 以及新生和成年小鼠心肌细胞。目前所有项目都在使用核心。《核心》 该机构现在是，也将由克里斯托弗·贝恩斯博士领导，他是在 2005年儿科分子心血管生物学教研室。贝恩斯博士有广泛的 心肌细胞制备和重组腺病毒载体的制备和使用的经验 心肌细胞生物学研究(Baines等人，2005年)。他在2005年担任核心运营的监督员 奥兰多·布埃诺医生离开的时候。必要的人员已经在实验室工作，并已充分 训练有素。Core C将提供各种支持服务，包括： A.生产穿梭载体质粒DNA(分发给研究人员) B.将转基因穿梭载体电穿孔导入含腺病毒基因组DNA的细菌 重组 C.阳性重组腺病毒的鉴定和纯化 D.维持允许腺病毒复制的人胚胎肾(HEK)293细胞 E.重组培养物的扩增 重组腺病毒的纯化和浓缩 G.用于确定斑块形成单位(PFU)水平的存量滴定 H.用于存储和编目低通道病毒库存的储存库服务，作为病毒的备份 提供给调查人员的股票 一、新生大鼠心肌细胞培养的周备 J.根据需要准备新生和成年小鼠心肌细胞培养

The Core will provide necessary support for the creation, selection, characterization, production, and purification of recombinant adenoviruses, as well as the isolation and culture of neonatal rat cardiomyocytes and neonatal and adult mouse cardiomyocytes. The Core is currently used by all of the Projects. The Core facility is and will be directed by Christopher Baines, Ph.D., who was appointed to the faculty in the Department of Pediatrics in 2005, Division of Molecular Cardiovascular Biology. Dr. Baines has extensive experience in cardiomyocyte preparation and in the generation and use of recombinant adenoviral vectors in the study of myocardial cell biology (Baines et al, 2005). He assumed oversight of Core operations in 2005 when Dr. Orlando Bueno left. The necessary personnel are already working in the laboratory and are fully trained. Core C will provide a variety of support services including: a. Production of the shuttle vector plasmid DNA (for distribution to investigators) b. Electroporation of transgene shuttle vector into adenoviral genomic DNA-containing bacteria for recombination c. Identification and purification of positive adenoviral recombinants d. Maintenance of human embryonic kidney (HEK) 293 cells permissive for adenoviral replication e. Amplification of recombinant cultures f. Purification and concentration of recombinant adenoviruses g. Titration of stocks for determination of plaque forming unit (pfu) levels h. Repository service for storage and cataloging of low passage viral stocks to serve as a backup for viral stocks given to investigators i. Weekly preparation of neonatal rat cardiomyocyte cultures j. Preparation of neonatal and adult mouse cardiomyocyte cultures when required