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用于药物发现的淋巴瘤患者来源的肿瘤异种移植模型的体内和体外研究

In Vivo and Ex Vivo Patient-Derived Tumor Xenograft Models of Lymphoma for Drug Discovery.

机构信息

Department of Pathology and Laboratory Medicine, Weill Cornell Medicine, New York, New York.

Department of Translational and Precision Medicine, Sapienza University of Rome, Rome, Italy.

出版信息

Curr Protoc. 2021 Apr;1(4):e96. doi: 10.1002/cpz1.96.

Abstract

In the hemato-oncology field, remarkable scientific progress has been achieved, primarily propelled by the discovery of new technologies, improvement in genomics, and novel in vitro and in vivo models. The establishment of multiple cell line collections and the development of instrumental mouse models enhanced our ability to discover effective therapeutics. However, cancer models that faithfully mimic individual cancers are still imperfect. Patient-derived tumor xenografts (PDTXs) have emerged as a powerful tool for identifying the mechanisms which drive tumorigenesis and for testing potential therapeutic interventions. The recognition that PDTXs can maintain many of the donor samples' properties enabled the development of new strategies for discovering and implementing therapies. Described in this article are protocols for the generation and characterization of lymphoma PDTXs that may be used as the basis of shared procedures. Universal protocols will foster the model utilization, enable the integration of public and private repositories, and aid in the development of shared platforms. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Tissue handling and cryopreservation of primary and PDTX samples Basic Protocol 2: Performing tumor implant in immunocompromised mice PDTX models Alternate Protocol 1: Intra-medullary femoral injection Alternate Protocol 2: Intravenous injection Alternate Protocol 3: Intraperitoneal injection Support Protocol 1: Phenotypical characterization of PDTXs by flow cytometry Support Protocol 2: Biological and molecular characterization of PDTX tumors by PCR detection of IGK, IGH, and TCR rearrangements Basic Protocol 3: Harvesting PDTX-derived tumor cells for ex vivo experiments Basic Protocol 4: In vivo testing of multiple compounds in a PDTX mouse model.

摘要

在血液肿瘤学领域,取得了显著的科学进展,主要得益于新技术的发现、基因组学的改进以及新型的体外和体内模型。多个细胞系集的建立和仪器化的小鼠模型的发展提高了我们发现有效治疗方法的能力。然而,能够真实模拟个体癌症的癌症模型仍然不够完善。患者来源的肿瘤异种移植(PDTX)已成为鉴定驱动肿瘤发生的机制和测试潜在治疗干预措施的有力工具。人们认识到 PDTX 可以保持供体样本的许多特性,这为发现和实施治疗策略提供了新的思路。本文描述了生成和鉴定淋巴瘤 PDTX 的方案,这些方案可以作为共享程序的基础。通用方案将促进模型的使用,实现公共和私人库的整合,并有助于开发共享平台。©2021Wiley Periodicals LLC. 基本方案 1:原发和 PDTX 样本的组织处理和冷冻保存 基本方案 2:在免疫功能低下的小鼠中进行肿瘤移植 PDTX 模型 可选方案 1:股骨髓内注射 可选方案 2:静脉注射 可选方案 3:腹腔内注射 支持方案 1:通过流式细胞术对 PDTX 进行表型特征分析 支持方案 2:通过 IGK、IGH 和 TCR 重排的 PCR 检测对 PDTX 肿瘤进行生物学和分子特征分析 基本方案 3:从 PDTX 衍生的肿瘤细胞中收获用于离体实验 基本方案 4:在 PDTX 小鼠模型中测试多种化合物的体内效果。

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