Simple-Flow:\nA 3D-Printed Multiwell Flow Plate to\nCoculture Primary Human Lung Cells at the Air–Liquid Interface

Immortalized epithelial cell lines and animal models\nhave been\nused in fundamental and preclinical research to study pulmonary diseases.\nHowever valuable, though, these models incompletely recapitulate the <i>in vivo</i> human lung, which leads to low predictive outcomes\nin potential respiratory treatments. Advanced technology and cell\nculture techniques stimulate the development of improved models that\nmore closely mimic the physiology of the human lung. Nonetheless,\nmost of these models are technically demanding and have a low throughput\nand reproducibility. Here, we describe a robust fluidic device consisting\nof a biocompatible and customizable 3D-printed cell culture plate,\nthe Simple-Flow, which has medium throughput, is simple to manufacture,\nand is easy to set up. As a proof of principle, human primary bronchial\nepithelial cells (hPBECs) and human pulmonary microvascular endothelial\ncells (hMVECs) were cocultured on the apical and basolateral sides\nof the inset membranes, respectively. While hPBECs were cultured at\nthe air–liquid interface to induce mucociliary differentiation,\nhMVECs were exposed to flow medium for up to 2 weeks. We show the\nversatility of 3D-printing technology in designing <i>in vitro</i> models for cell culturing applications, such as pediatric lung diseases\nor other pulmonary disorders.