Core C: CODEX Core

摘要(核心C) 我们计划的一个中心焦点是了解特定细胞类型的作用，包括免疫细胞 在PDAC在小鼠模型和人类中的发起、发展和传播的整个过程中。一个 这项工作的关键方面是定义 并了解这些参数如何随时间和对治疗的反应而变化。此外，虽然 每个项目都侧重于PDAC的不同方面，全面了解上皮细胞的作用 PDAC中的间隔和免疫系统需要同时分析这些细胞类型中的每一种 多个组织。为了实现这些目标，我们将使用索引联合检测(CODEX)这一新方法 由斯坦福大学诺兰博士的研究小组产生。Codex将允许对数十个组织切片进行细胞成像 抗体。然后，该核心中的食典数据可与个别项目中使用的CyTOF和scRNASeq中的补充数据相关联。核心C用于人类和小鼠胰腺研究的具体目标是： 1.为调查人员提供食典处理，包括染色、成像和质量控制 2.提供Codex分析，包括抗原聚类、细胞类型注释和邻域映射 3.与P01调查人员协商，为食典开发新的抗体和试剂 4.合作制定纸巾采购、加工、储存和检索的标准工作流程 与人类胰腺组织核心(核心B)。 项目1、2和3将专门使用食典核心平台来研究小鼠和人体组织。 核心C将与人类组织核心B密切合作，以确保适当的人体组织准备 支持食典分析。因此，Core C将提供技术上有难度且在大多数情况下无法提供的服务 实验室，商业上无法获得或无法在其他地方获得的材料，以及更可靠的服务 而且比在个别研究人员的实验室中进行的情况更具成本效益。这是法典的核心 将是新的，在斯坦福大学是独一无二的。

ABSTRACT (Core C) A central focus of our program is understanding the roles of specific cell types, including immune cell populations, throughout the initiation, development and spread of PDAC in mouse models and in humans. A key aspect of this effort is to define the frequency, spatial relationships and activation states of cell types within tumors, and to learn how these parameters change over time and in response to therapy. Additionally, while each project focuses on a different aspect of PDAC, a comprehensive understanding of the role of the epithelial compartment and the immune system in PDAC will require simultaneous analysis of each of these cell types in multiple tissues. To achieve these goals, we will use CO-Detection by indEXing (CODEX), a new method generated by Dr. Nolan’s group at Stanford. CODEX will allow cytometric imaging of tissue sections with dozens of antibodies. CODEX data from this Core can then be linked to complementary data from CyTOF and scRNASeq used in individual projects. The Specific Aims of Core C for human and mouse pancreas studies are: 1. Provide CODEX processing to investigators, including staining, imaging and quality control 2. Provide CODEX analysis, including antigen clustering, cell type annotation and neighborhood mapping 3. Develop new antibodies and reagents for CODEX in consultation with P01 investigators 4. Develop standard workflows for tissue procurement, processing, storage and retrieval in partnership with the human pancreas tissue core (Core B). Projects 1, 2 and 3 will specifically use the CODEX core platform for studies of both mouse and human tissues. Core C will work closely with the Human Tissue Core B to ensure appropriate human tissue preparation to support CODEX analysis. Thus, Core C will provide services technically difficult and not available in most laboratories, materials not available commercially or impossible to obtain elsewhere, and services more reliably and cost-effectively performed than if performed in an individual investigator’s laboratory. This CODEX Core would be new, and unique at Stanford.