Oatway Christina, Hirsch Calley L, Gregorieff Alex
Lunenfeld-Tanenbaum Research Institute, Sinai Health System, Toronto, Canada.
Department of Pathology, McGill University, Montreal, Canada.
Methods Mol Biol. 2018;1765:315-331. doi: 10.1007/978-1-4939-7765-9_21.
The development of intestinal organoid technology has greatly accelerated research in the field of colorectal cancer. Contrary to traditional cancer cell lines, organoids are composed of multiple cell types arranged in 3D structures highly reminiscent of their native tissues. Thus, organoids provide a near-physiological and readily accessible model to study tissue morphogenesis, adult stem cell behavior and tumorigenesis. Here, we provide protocols for establishing intestinal organoid cultures from genetically modified mouse lines and describe methods to overexpress and knockout genes of interest using lentiviral-based approaches.
肠道类器官技术的发展极大地加速了结直肠癌领域的研究。与传统癌细胞系不同,类器官由多种细胞类型组成,排列成高度类似于其天然组织的三维结构。因此,类器官为研究组织形态发生、成体干细胞行为和肿瘤发生提供了一个近乎生理且易于获取的模型。在此,我们提供了从基因修饰小鼠品系建立肠道类器官培养物的方案,并描述了使用基于慢病毒的方法过表达和敲除感兴趣基因的方法。