IMAGING ONCOLYTIC ADENOVIRUSES SPREAD IN A 3D SYSTEM

对 3D 系统中传播的溶瘤腺病毒进行成像

• 批准号: 8365770
• 负责人: HUNG Y. FAN
• 金额: $ 0.55万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-20 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8365770
• 关键词: Adenoviruses; Appearance; Biological Models; Biology; Cells; Data; Fluorescence; Funding; Grant; Image; Individual; Infection; Laboratories; Microscope; Monitor; National Center for Research Resources; Oncolytic; Optics; Principal Investigator; Research; Research Infrastructure; Resource Sharing; Resources; Solid; Source; Suspension substance; Suspensions; System; United States National Institutes of Health; Viral; Virus; cost; matrigel; monolayer; reconstitution; research study; three dimensional structure

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Study of viral spread in 3D cultures. The most relevant system for modeling viral spread would be in solid cell masses. As shown in the preliminary data, we have been able to image individual infected cells in solid masses by reconstituting infected and uninfected cells into spheres on Matrigel. We will first carry out analysis on AdEGFPuci-infected 293 cells. We will first infect a monolayer of 293 cells with AdEGFPuci, and 6 hr after infection, the infected cells will be trypsinized. The infected single cell suspensions will be mixed with an excess of uninfected 293 cells, and plated onto Matrigel matrices to allow formation of cell aggregates and spheres. The spheres will be monitored daily for appearance of green fluorescent cells and fluorescent Z-stack montages will be recorded and compiled for any sphere that initially show one infected cell. A confocal microscope suitable for these experiments is available in the campus optical biology shared resource. It seems possible that the rate of virus spread may be more rapid in 3D structures than in monolayer, or that the number of secondarily infected cells may be larger. It will also be possible to determine the effects of initial MOI on spread through 3D cultures.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 研究在3D培养物中的病毒扩散。建模病毒扩散的最相关系统将是固体细胞质量。如初步数据所示，我们已经能够通过将受感染和未感染的细胞重构为基质中的球体来对固体中的单个感染细胞进行成像。 我们将首先对ADEGFPUCI感染的293个细胞进行分析。我们将首先感染具有ADEGFPUCI的293个细胞的单层，感染后6小时，感染的细胞将被胰蛋白酶粉碎。感染的单细胞悬浮液将与过量未感染的293个细胞混合，并将其镀到矩阵矩阵上，以形成细胞聚集体和球体。每天将监视这些球体，以显示绿色荧光细胞的出现，并将记录并为任何最初显示一个受感染细胞的球体记录并编译。校园光学生物学共享资源可提供适合这些实验的共聚焦显微镜。在3D结构中，病毒扩散率似乎比单层中可能更快，或者次要感染细胞的数量可能更大。还可以确定初始MOI对通过3D培养物扩散的影响。