Cell Culture Core

细胞培养核心将位于弗吉尼亚大学卫生系统医学研究大楼4的2楼，这是肺S-硝酸化信号研究中心的中心位置。该设施距离医院主手术室约100码，距离 尸检室，毗邻Palmer博士使用的隔室(项目2)，高于Lewis博士使用的隔室的两层(项目3)。该设施将配备4个指定的细胞培养孵化器、两个冰箱、两个显微镜、一个指定的组织培养罩、解剖显微镜、来自Fine Science Tools(加利福尼亚州福斯特市)的解剖设备、专门用于细胞培养的微型离心机、两个-20°C冷冻机、一个-80°C冰柜以及一个用于细胞系的液氮储存库。所有用于细胞培养的试剂都将由核心提供。细胞培养核心设施将设在儿科组织学核心设施的隔壁，该设施完全能够进行冷冻和石蜡包埋 切片和免疫组织化学染色。这与加斯顿博士的组织培养设施相邻，在那里将进行循环压应力。细胞培养核心的人员在呼吸道上皮细胞、内皮细胞和原代神经细胞培养工作的各个方面都拥有广泛的专业知识。通过北卡罗来纳大学的兰德尔博士和外科的琼斯博士的合作，将为实验提供CF和非CF肺组织。来自弗吉尼亚大学病理学系的Moskaluk博士将帮助保存和收获我们的CF和非CF肺组织，以及协调供应通知。Kelly，RN，是护士协调员，将帮助我们识别患者的性别、年龄和吸烟状况，并获得受试者法定监护人的知情同意。此外，凯利将确保遵守机构审查委员会的要求。

The Cell Culture Core will be located on the 2rd floor of medical research building 4 at the University of Virginia Health System, a central location within the Pulmonary S-Nitrosylation Signaling Research Center. This facility is located approximately 100 yards from the main hospital operating room, approximately 150 yards from the autopsy suite, next to the building housing the vivarium used by Dr. Palmer (Project 2) and two floors above the vivarium used by Dr. Lewis (Project 3). This facility will be equipped with 4 designated cell culture incubators, two refrigerators, two microscopes, a designated tissue culture hood, dissecting microscopes, dissecting equipment from Fine Science Tools (Foster City, California), microcentrifuges exclusively for cell culture, two -20¿C freezers, one -80¿C freezer as well as a liquid nitrogen storage for the cell lines. All reagents for cell cultures will be provided by the core. The cell culture core facility will be located next door to the Pediatric Histology Core Facility which is fully capable of performing frozen and paraffin-embedded sectioning and immunohistochemistry. This is adjacent to Dr. Gaston's tissue culture facility in which cyclic compressive stress will be performed. Personnel in the Cell Culture Core have extensive expertise in all aspects of airway epithelial cell, endothelial and primary neuronal cell culture work. Collaborations through Dr. Randell at the University of North Carolina and Dr. Jones in the Department of Surgery will provide CF and non-CF lung tissue for experiments. Dr. Moskaluk, from the University of Virginia Department of Pathology, will help to preserve and harvest our CF and non-CF lung tissues, as well as coordinating notification of availability. Kelly, RN, is the nurse-coordinator will help us to identify gender, age, and smoking status of patients as well as obtain informed consent from the legal guardian of subjects. In addition, Ms. Kelly will assure compliance with Institutional Review Board requirements.