A Three-Dimensional Organoid Culture System Derived from Human Glioblastomas Recapitulates the Hypoxic Gradients and Cancer Stem Cell Heterogeneity of Tumors Found In Vivo.
A Three-Dimensional Organoid Culture System Derived from Human Glioblastomas Recapitulates the Hypoxic Gradients and Cancer Stem Cell Heterogeneity of Tumors Found In Vivo.
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DOI：
10.1158/0008-5472.can-15-2402
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发表时间：
2016-04-15
期刊：
影响因子：
11.2
通讯作者：
Rich JN
中科院分区：
文献类型：
--
作者：
Hubert CG;Rivera M;Spangler LC;Wu Q;Mack SC;Prager BC;Couce M;McLendon RE;Sloan AE;Rich JN
许多癌症的特征是由肿瘤起始的癌症干细胞（CSC）驱动的细胞层级，并依赖于与肿瘤微环境的复杂相互作用。标准细胞培养条件不能重现原始肿瘤结构或微环境梯度，并且未设计为保留亲本肿瘤的细胞异质性。在这里，我们描述了一种三维培养系统，该系统支持直接来源于胶质母细胞瘤标本的肿瘤类器官的长期生长和扩增，包括患者来源的原代培养物、异种移植物、基因工程胶质瘤模型或患者样本。来源于患者肿瘤多个区域的类器官保留选择性致瘤潜力。此外，类器官可以直接从通常不适合体外培养的脑转移瘤中建立。一旦形成，肿瘤类器官就会生长数月，并表现出区域异质性，快速分裂的SOX 2+、OLIG 2+和TLX+细胞外部区域围绕着主要非干细胞衰老细胞和弥漫性静止CSC的缺氧核心。值得注意的是，类器官内的非干细胞对放射治疗敏感，而相邻的CSC具有放射抗性。患者源性类器官的原位移植导致肿瘤显示出组织学特征，包括单细胞侵袭性，与患者源性球体培养物形成的肿瘤相比，其更能代表亲本肿瘤。总之，我们提出了一种新的离体模型，其中表型多样的干细胞和非干细胞胶质母细胞瘤细胞群可以同时培养，以探索微环境影响和CSC生物学的新方面。
Many cancers feature cellular hierarchies that are driven by tumor-initiating, cancer stem cells (CSCs) and rely on complex interactions with the tumor microenvironment. Standard cell culture conditions fail to recapitulate the original tumor architecture or microenvironmental gradients, and are not designed to retain the cellular heterogeneity of parental tumors. Here, we describe a three-dimensional culture system that supports the long-term growth and expansion of tumor organoids derived directly from glioblastoma specimens, including patient-derived primary cultures, xenografts, genetically engineered glioma models, or patient samples. Organoids derived from multiple regions of patient tumors retain selective tumorigenic potential. Furthermore, organoids could be established directly from brain metastases not typically amenable to in vitro culture. Once formed, tumor organoids grew for months and displayed regional heterogeneity with a rapidly dividing outer region of SOX2+, OLIG2+, and TLX+ cells surrounding a hypoxic core of primarily non-stem senescent cells and diffuse, quiescent CSCs. Notably, non-stem cells within organoids were sensitive to radiation therapy, whereas adjacent CSCs were radioresistant. Orthotopic transplantation of patient-derived organoids resulted in tumors displayed histological features, including single cell invasiveness, that were more representative of the parental tumor compared with those formed from patient-derived sphere cultures. In conclusion, we present a new ex vivo model in which phenotypically diverse stem and non-stem glioblastoma cell populations can be simultaneously cultured to explore new facets of microenvironmental influences and CSC biology.