Center for Regenerative Medicine of Boston University and Boston Medical Center, Boston, MA 02118, USA; The Pulmonary Center and Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA.
Center for Regenerative Medicine of Boston University and Boston Medical Center, Boston, MA 02118, USA.
Stem Cell Reports. 2018 May 8;10(5):1579-1595. doi: 10.1016/j.stemcr.2018.03.013. Epub 2018 Apr 12.
Lung epithelial lineages have been difficult to maintain in pure form in vitro, and lineage-specific reporters have proven invaluable for monitoring their emergence from cultured pluripotent stem cells (PSCs). However, reporter constructs for tracking proximal airway lineages generated from PSCs have not been previously available, limiting the characterization of these cells. Here, we engineer mouse and human PSC lines carrying airway secretory lineage reporters that facilitate the tracking, purification, and profiling of this lung subtype. Through bulk and single-cell-based global transcriptomic profiling, we find PSC-derived airway secretory cells are susceptible to phenotypic plasticity exemplified by the tendency to co-express both a proximal airway secretory program as well as an alveolar type 2 cell program, which can be minimized by inhibiting endogenous Wnt signaling. Our results provide global profiles of engineered lung cell fates, a guide for improving their directed differentiation, and a human model of the developing airway.
肺上皮谱系在体外很难以纯形式维持，谱系特异性报告基因已被证明对于监测其从培养的多能干细胞 (PSC) 中出现非常有价值。然而，以前没有用于跟踪源自 PSC 的近端气道谱系的报告基因构建体，限制了对这些细胞的表征。在这里，我们构建了携带气道分泌谱系报告基因的小鼠和人类 PSC 系，这些报告基因有助于这些肺亚型的跟踪、纯化和分析。通过批量和单细胞全转录组谱分析，我们发现 PSC 衍生的气道分泌细胞易受表型可塑性的影响，例如倾向于同时表达近端气道分泌程序和肺泡 2 型细胞程序，通过抑制内源性 Wnt 信号可以最小化这种情况。我们的研究结果提供了工程化肺细胞命运的全局图谱，为改善其定向分化提供了指导，并为发育中的气道提供了人类模型。
