生成用于共聚焦延时成像和空间分析整合的3D体外小鼠和患者来源的胶质瘤外植体切片模型。

Generation of 3D ex vivo mouse- and patient-derived glioma explant slice model for integration of confocal time-lapse imaging and spatial analysis.

作者信息

Comba Andrea, Varela Maria Luisa, Faisal Syed M, Abel Clifford C, Argento Anna E, Al-Holou Wajd N, Hollon Todd C, Perelman Jacqueline D, Dunn Patrick J, Motsch Sebastien, Castro Maria G, Lowenstein Pedro R

机构信息

Department of Neurosurgery, University of Michigan Medical School, Ann Arbor, MI 48109, USA; Department of Cell and Developmental Biology, University of Michigan Medical School, Ann Arbor, MI 48109, USA; Rogel Cancer Center, University of Michigan Medical School, Ann Arbor, MI 48109, USA.

出版信息

STAR Protoc. 2023 Mar 16;4(2):102174. doi: 10.1016/j.xpro.2023.102174.

DOI:10.1016/j.xpro.2023.102174

PMID:36930648

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10036861/

Abstract

Development of spatial-integrative pre-clinical models is needed for glioblastoma, which are heterogenous tumors with poor prognosis. Here, we present an optimized protocol to generate three-dimensional ex vivo explant slice glioma model from orthotopic tumors, genetically engineered mouse models, and fresh patient-derived specimens. We describe a step-by-step workflow for tissue acquisition, dissection, and sectioning of 300-μm tumor slices maintaining cell viability. The explant slice model allows the integration of confocal time-lapse imaging with spatial analysis for studying migration, invasion, and tumor microenvironment, making it a valuable platform for testing effective treatment modalities. For complete details on the use and execution of this protocol, please refer to Comba et al. (2022)..

摘要

胶质母细胞瘤需要开发空间整合的临床前模型，胶质母细胞瘤是预后不良的异质性肿瘤。在此，我们提出了一种优化方案，用于从原位肿瘤、基因工程小鼠模型和新鲜的患者来源标本中生成三维体外外植体切片胶质瘤模型。我们描述了一个组织获取、解剖和切片300μm肿瘤切片并保持细胞活力的分步工作流程。外植体切片模型允许将共聚焦延时成像与空间分析相结合，以研究迁移、侵袭和肿瘤微环境，使其成为测试有效治疗方式的宝贵平台。有关本方案使用和执行的完整详细信息，请参阅Comba等人（2022年）。