Protocol for the derivation and alveolar type 2 differentiation of late-stage lung tip progenitors from the developing human lungs

Abstract

Molecular and cellular mechanisms of human lung alveolar development are poorly understood due to a lack of in vitro model systems. This protocol details the isolation, derivation, and genetic modification of lung tip epithelial progenitors from human fetal lungs. It includes steps for isolating distal lung epithelial cells, expanding tip progenitor organoids, culturing tip organoids in vitro, and differentiating them into alveolar type 2 cells. This will aid in understanding alveolar differentiation mechanisms and neonatal diseases. For complete details on the use and execution of this protocol, please refer to Lim et al.¹.

Keywords: Cell Differentiation; Cell culture; Cell isolation; Cell separation/fractionation; Developmental biology; Flow Cytometry; Organoids.