A surgical orthotopic organoid transplantation approach in mice to visualize and study colorectal cancer progression
A surgical orthotopic organoid transplantation approach in mice to visualize and study colorectal cancer progression
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DOI：
10.1038/nprot.2017.137
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发表时间：
2018-02-01
期刊：
影响因子：
14.8
通讯作者：
Drost, Jarno
中科院分区：
文献类型：
--
作者：
Fumagalli, Arianna;Suijkerbuijk, Saskia J. E.;Drost, Jarno
目前大多数可用的结直肠癌（CRC）小鼠模型不适合研究转移阶段的进展。最近，来自鼠CRC模型或患者的肿瘤类器官系的建立，以及用基因组编辑技术对其进行工程改造的可能性，已经提供了大量忠实地再现天然肿瘤的表型和遗传异质性的肿瘤材料。为了研究肿瘤在自然体内环境中的进展，我们开发了一种基于将CRC类器官移植到盲肠上皮中的原位方法。这里详细描述了20分钟的过程，并且使移植的类器官能够在肠道内生长成单个肿瘤块。由于潜伏期长，肿瘤细胞能够通过血液循环扩散并在远处部位形成转移。该方法被设计用于产生适合于研究CRC进展的肿瘤，从而提供通过活体显微镜术在体内真实的时间可视化肿瘤细胞动力学的机会。
Most currently available colorectal cancer (CRC) mouse models are not suitable for studying progression toward the metastatic stage. Recently, establishment of tumor organoid lines, either from murine CRC models or patients, and the possibility of engineering them with genome- editing technologies, have provided a large collection of tumor material faithfully recapitulating phenotypic and genetic heterogeneity of native tumors. To study tumor progression in the natural in vivo environment, we developed an orthotopic approach based on transplantation of CRC organoids into the cecal epithelium. The 20-min procedure is described in detail here and enables growth of transplanted organoids into a single tumor mass within the intestinal tract. Due to long latency, tumor cells are capable of spreading through the blood circulation and forming metastases at distant sites. This method is designed to generate tumors suitable for studying CRC progression, thereby providing the opportunity to visualize tumor cell dynamics in vivo in real time by intravital microscopy.