In vitro generation of human pluripotent stem cell derived lung organoids

TLDR

3D organoid cultures have created physiologically complex in vitro models for studying human development and disease. The study reports a stepwise differentiation protocol converting human pluripotent stem cells into lung organoids. The protocol manipulates developmental signaling to generate ventral‑anterior foregut spheroids that expand into human lung organoids. The resulting HLOs contain epithelial and mesenchymal compartments resembling native lung, including airway‑like epithelium with basal and immature ciliated cells, smooth muscle and myofibroblasts, an alveolar‑like domain, and transcriptomic profiles that closely match human fetal lung.

Abstract

Recent breakthroughs in 3-dimensional (3D) organoid cultures for many organ systems have led to new physiologically complex in vitro models to study human development and disease. Here, we report the step-wise differentiation of human pluripotent stem cells (hPSCs) (embryonic and induced) into lung organoids. By manipulating developmental signaling pathways hPSCs generate ventral-anterior foregut spheroids, which are then expanded into human lung organoids (HLOs). HLOs consist of epithelial and mesenchymal compartments of the lung, organized with structural features similar to the native lung. HLOs possess upper airway-like epithelium with basal cells and immature ciliated cells surrounded by smooth muscle and myofibroblasts as well as an alveolar-like domain with appropriate cell types. Using RNA-sequencing, we show that HLOs are remarkably similar to human fetal lung based on global transcriptional profiles, suggesting that HLOs are an excellent model to study human lung development, maturation and disease.