Department of Cell and Developmental Biology, University of Michigan Medical School, Ann Arbor, MI 48109, USA.
Department of Computational Medicine and Bioinformatics, University of Michigan Medical School, Ann Arbor, MI 48109, USA.
Development. 2022 Oct 15;149(20). doi: 10.1242/dev.200693. Epub 2022 Aug 30.
Bud tip progenitors (BTPs) in the developing lung give rise to all epithelial cell types found in the airways and alveoli. This work aimed to develop an iPSC organoid model enriched with NKX2-1+ BTP-like cells. Building on previous studies, we optimized a directed differentiation paradigm to generate spheroids with more robust NKX2-1 expression. Spheroids were expanded into organoids that possessed NKX2-1+/CPM+ BTP-like cells, which increased in number over time. Single cell RNA-sequencing analysis revealed a high degree of transcriptional similarity between induced BTPs (iBTPs) and in vivo BTPs. Using FACS, iBTPs were purified and expanded as induced bud tip progenitor organoids (iBTOs), which maintained an enriched population of bud tip progenitors. When iBTOs were directed to differentiate into airway or alveolar cell types using well-established methods, they gave rise to organoids composed of organized airway or alveolar epithelium, respectively. Collectively, iBTOs are transcriptionally and functionally similar to in vivo BTPs, providing an important model for studying human lung development and differentiation.
芽基祖细胞（BTPs）在发育中的肺中产生所有在气道和肺泡中发现的上皮细胞类型。这项工作旨在开发一种富含 NKX2-1+BTP 样细胞的 iPSC 类器官模型。在前人的研究基础上，我们优化了定向分化方案，以生成具有更强 NKX2-1 表达的球体。球体被扩展成具有 NKX2-1+/CPM+BTP 样细胞的类器官，这些细胞的数量随时间增加。单细胞 RNA 测序分析显示，诱导的 BTP（iBTP）与体内 BTP 之间具有高度的转录相似性。使用 FACS，iBTP 被纯化并作为诱导芽基祖细胞类器官（iBTO）扩增，这些类器官保持了芽基祖细胞的富集群体。当使用成熟的方法将 iBTO 定向分化为气道或肺泡细胞类型时，它们分别产生了由组织化气道或肺泡上皮组成的类器官。总的来说，iBTO 在转录和功能上与体内 BTP 相似，为研究人类肺发育和分化提供了重要模型。
