目前，器官样培养是不同器官进行体外研究的重要工具。小鼠小肠隐窝可以形成器官样体，当在3D胞外基质中培养时，这种器官样体可以模拟肠上皮。器官样体由各种细胞类型组成，这些细胞类型完成各种肠道稳态功能。其中包括泛特细胞、肠内分泌细胞、肠上皮细胞、高尔基细胞和簇状细胞。将经充分确认的分子添加到培养基中以富集具有富含G蛋白偶联受体5的富勒氨酸重复的肠道干细胞（ISCs），并用于将分化驱动到特定细胞系；这些分子包括表皮生长因子、Noggin（一种骨形态发生蛋白）和R-spondin 1。此外，还详细介绍了通过单个促红细胞生成素产生的肝细胞受体B2（EphB2）阳性ISC生成器官样体的方案。在该方法文章中，描述了从组织中分离小肠隐窝和单个ISC以及确保高效建立器官样体的技术。

At present, organoid culture represents an important tool for in vitro studies of different biological aspects and diseases in different organs. Murine small intestinal crypts can form organoids that mimic the intestinal epithelium when cultured in a 3D extracellular matrix. The organoids are composed of all cell types that fulfill various intestinal homeostatic functions. These include Paneth cells, enteroendocrine cells, enterocytes, goblet cells, and tuft cells. Well-characterized molecules are added into the culture medium to enrich the intestinal stem cells (ISCs) labeled with leucine-rich repeats containing G protein-coupled receptor 5 and are used to drive differentiation down specific lineages; these molecules include epidermal growth factor, Noggin (a bone morphogenetic protein), and R-spondin 1. Additionally, a protocol to generate organoids from a single erythropoietin-producing hepatocellular receptor B2 (EphB2)-positive ISC is also detailed. In this methods article, techniques to isolate small intestinal crypts and a single ISC from tissues and ensure the efficient establishment of organoids are described.