Reconstituted basement membrane enables airway epithelium modeling and nanoparticle toxicity testing

Basement membrane (BM) acts as a sheet-like extracellular matrix to support and promote the formation of epithelial and endothelial cell layers. The in vitro reconstruction of the BM is however not easy due to its ultrathin membrane features. This difficulty is overcome by self-assembling type IV collagen and laminin in the porous areas of a monolayer of crosslinked gelatin nanofibers deposited on a honeycomb microframe. Herein, a method is presented to generate airway epithelium by using such an artificial basement membrane (ABM) and human-induced pluripotent stem cells (hiPSCs). Bipolar primordial lung progenitors are firstly induced from hiPSCs and then replated on the ABM for differentiation toward matured airway epithelium under submerged and air-liquid interface culture conditions. As a result, a pseudostratified airway epithelium consisting of several cell types is achieved, showing remarkable apical secretion of MUC5AC proteins and clear advantages over other types of substrates. As a proof of concept, the derived epithelium is used for toxicity test of cadmium telluride (CdTe) nanoparticles (NPs), demonstrating the applicability of ABM-based assays involving hiPSC-derived epithelial cells-based assays.