A Protocol for Rapid Post-mortem Cell Culture of Diffuse Intrinsic Pontine Glioma (DIPG).
A Protocol for Rapid Post-mortem Cell Culture of Diffuse Intrinsic Pontine Glioma (DIPG).
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DOI：
10.3791/55360
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发表时间：
2017-03-07
期刊：
影响因子：
--
通讯作者：
Monje M
中科院分区：
文献类型：
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作者：
Lin GL;Monje M
弥漫性内在脑桥胶质瘤（DIPG）是一种儿童脑干肿瘤，具有普遍致命的预后。由于手术切除不是可行的治疗策略，并且不定期进行活检，因此用于研究的患者样本的可用性有限。因此，研究这种疾病的努力受到缺乏可靠疾病模型的挑战。为了满足这一需求，我们在这里描述了一个快速处理死后尸检组织样本的方案，以产生持久的患者来源的细胞培养模型，可用于体外试验或体内原位异种移植实验。这些模型可用于筛选潜在的药物靶点并研究DIPG中的基本病理生物学过程。该方案可以进一步扩展到使用荧光激活细胞分选（FACS）分析和分离肿瘤和微环境细胞，这使得能够在大量细胞或单细胞水平上对基因表达、蛋白质表达或DNA的表观遗传修饰进行后续分析。最后，该方案也可以适用于产生其他中枢神经系统肿瘤的患者来源的培养物。
Diffuse Intrinsic Pontine Glioma (DIPG) is a childhood brainstem tumor that carries a universally fatal prognosis. Because surgical resection is not a viable treatment strategy and biopsy is not routinely performed, the availability of patient samples for research is limited. Consequently, efforts to study this disease have been challenged by a paucity of faithful disease models. To address this need, we describe here a protocol for the rapid processing of post-mortem autopsy tissue samples in order to generate durable patient-derived cell culture models that can be used in in vitro assays or in vivo orthotopic xenograft experiments. These models can be used to screen for potential drug targets and to study fundamental pathobiological processes within DIPG. This protocol can further be extended to analyze and isolate tumor and microenvironmental cells using Fluorescence-activated Cell Sorting (FACS), which enables subsequent analysis of gene expression, protein expression, or epigenetic modifications of DNA at the bulk cell or single cell level. Finally, this protocol can also be adapted to generate patient-derived cultures for other central nervous system tumors.