肠道类器官3D和2D培养及组织学分析的更新方案
Updated Protocols for 3D and 2D Culture and Histological Analysis of Intestinal Organoids.
摘要(中文译文)
肠上皮通过Wnt、骨形态发生蛋白、表皮生长因子（EGF）和Notch等多个信号通路的整合调控，维持持续自我更新和多向分化能力。基于这一原理，表皮生长因子、Noggin和Wnt激动剂R-spondin的确定组合已被证明能够支持小鼠肠道干细胞的长期扩增，并形成具有无限自我更新和完全分化能力的类器官。在人肠道类器官培养中，添加靶向p38和TGF-β通路的小分子抑制剂可促进上皮增殖，而用胰岛素样生长因子-1和成纤维细胞生长因子-2替代表皮生长因子和p38抑制剂则能够实现高效的多谱系分化。近年来，肠道类器官的单层培养因其可直接接触顶端膜以及与肠道微生物群共培养的优势而日益得到应用。在此，我们总结近期在人肠道类器官培养和组织学分析方面的技术进展，提供更新的实验方案，以加深对肠道稳态和肠道疾病发病机制的理解。
Abstract
The intestinal epithelium maintains continuous self-renewal and multi-differentiation capacities through the integrated regulation of multiple signaling pathways, including Wnt, bone morphogenetic protein, epidermal growth factor (EGF), and Notch signaling pathways. Based on this principle, a defined combination of EGF, Noggin, and the Wnt agonist R-spondin has been shown to support long-term expansion of mouse intestinal stem cells and the formation of organoids with indefinite self-renewal and full differentiation capacity. In human intestinal organoid cultures, the addition of small molecule inhibitors targeting p38 and TGF-β pathways facilitated epithelial propagation, while replacement of EGF and a p38 inhibitor with insulin-like growth factor-1 and fibroblast growth factor-2 enabled efficient multilineage differentiation. Recently, the monolayer culture of intestinal organoid has been increasingly used due to its advantages of direct access to the apical membrane and its ability to co-culture with gut microbiota. Here, we summarize our recent technological advances in human intestinal organoid culture and histological analysis, providing updated protocols that will deepen the understanding of intestinal homeostasis and the pathogenesis of intestinal diseases.
如何引用
AMA
Hikaru Hanyu, Shinya Sugimoto, Toshiro Sato. Updated Protocols for 3D and 2D Culture and Histological Analysis of Intestinal Organoids.. Methods in molecular biology (Clifton, N.J.). 2027; doi:10.1007/978-1-0716-5412-5_9.
APA
Hikaru Hanyu, Shinya Sugimoto, Toshiro Sato (2027). Updated Protocols for 3D and 2D Culture and Histological Analysis of Intestinal Organoids.. Methods in molecular biology (Clifton, N.J.). https://doi.org/10.1007/978-1-0716-5412-5_9
GB/T 7714
Hikaru Hanyu, Shinya Sugimoto, Toshiro Sato. Updated Protocols for 3D and 2D Culture and Histological Analysis of Intestinal Organoids.[J]. Methods in molecular biology (Clifton, N.J.), 2027 doi:10.1007/978-1-0716-5412-5_9.
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