Stem Cell Institute, Ankara University, Ankara, Turkey.
Faculty of Medicine, Department of Biophysics, Ankara University, Ankara, Turkey.
Methods Mol Biol. 2024;2835:59-67. doi: 10.1007/978-1-0716-3995-5_6.
Organoids, three-dimensional, stem cell-based structures that mimic the cellular and functional architecture of tissues, have emerged as an innovative in vitro tool. They offer highly efficient models for studying both embryonic development and disease progression processes. Colon organoids can also be generated from biopsies obtained during a colonoscopy. However, the invasive nature of biopsy collection poses practical challenges and introduces biases when studying patients who are already afflicted. Therefore, the use of iPSC-derived colon organoids can be considered a more practical approach for researchers and patients alike. Numerous protocols have been published for generating colon organoids from iPSCs. While most of these protocols share a common developmental process, some are labor-intensive or require additional equipment. Taking these considerations into account, we present a cost-effective and straightforward yet functionally robust colon organoid protocol: (1) definitive endoderm differentiation, (2) hindgut endoderm differentiation, and (3) maturation of colon spheroids into mature organoids.
类器官是一种基于干细胞的三维结构，可模拟组织的细胞和功能结构，已成为一种创新的体外工具。它们为研究胚胎发育和疾病进展过程提供了高效的模型。也可以从结肠镜检查中获得的活检中生成结肠类器官。然而，活检采集的侵入性性质在研究已经患病的患者时带来了实际挑战和偏差。因此，使用 iPSC 衍生的结肠类器官可以被认为是研究人员和患者都更实用的方法。已经发表了许多从 iPSC 生成结肠类器官的方案。虽然大多数方案都有一个共同的发育过程，但有些方案劳动强度大或需要额外的设备。考虑到这些因素，我们提出了一种具有成本效益、简单而又功能强大的结肠类器官方案：（1）确定内胚层分化，（2）后肠内胚层分化，以及（3）结肠球体成熟为成熟的类器官。
