Deniz Derman I,
High-throughput bioprinting of the nasal epithelium using patient-derived nasal epithelial cells. Biofabrication. 2023;15(4):044103.
JGM, Humans, Bioprinting, Nasal Mucosa, Epithelial Cells, Respiratory Mucosa, Cilia
This research was primarily supported by the National Institute of Health Award #U19AI142733. We gratefully acknowledge the contribution of the Single Cell Biology service, the Genome Technologies service, and cyberinfrastructure high performance computing resources at The Jackson Laboratory (JAX) for expert assistance with the work described herein. These shared services were supported in part by the JAX Cancer Center (P30 CA034196). J O was additionally supported by 1DP2GM126893-01, 1 R01 AR078634-01 and 5 R21 AR075174-02. M H was sup- ported by T32HG010463. The Cure Cystic Fibrosis Columbus (C3) Epithelial Cell Core at Nationwide Children’s Hospital (NCH) provided primary human nasal epithelial cultures for this work, with the help of the NCH Biopathology Center Core and Data Collaboration Team. C3 is supported by a Cystic Fibrosis Foundation (CFF) Research Development Program Grant (MCCOY17R2) and a CFF grant to the NCH Division of Pediatric Pulmonary Medicine (MCCOY19RO). The authors are also thankful to Dr Adolfo García-Sastre and Michael Schotsaert from Icahn School of Medicine at Mount Sinai, New York, NY for providing, GFP+ PR8 virus.
Progenitor human nasal epithelial cells (hNECs) are an essential cell source for the reconstruction of the respiratory pseudostratified columnar epithelium composed of multiple cell types in the context of infection studies and disease modeling. Hitherto, manual seeding has been the dominant method for creating nasal epithelial tissue models through biofabrication. However, this approach has limitations in terms of achieving the intricate three-dimensional (3D) structure of the natural nasal epithelium. 3D bioprinting has been utilized to reconstruct various epithelial tissue models, such as cutaneous, intestinal, alveolar, and bronchial epithelium, but there has been no attempt to use of 3D bioprinting technologies for reconstruction of the nasal epithelium. In this study, for the first time, we demonstrate the reconstruction of the nasal epithelium with the use of primary hNECs deposited on Transwell inserts via droplet-based bioprinting (DBB), which enabled high-throughput fabrication of the nasal epithelium in Transwell inserts of 24-well plates. DBB of progenitor hNECs ranging from one-tenth to one-half of the cell seeding density employed during the conventional cell seeding approach enabled a high degree of differentiation with the presence of cilia and tight-junctions over a 4 weeks air-liquid interface culture. Single cell RNA sequencing of these cultures identified five major epithelial cells populations, including basal, suprabasal, goblet, club, and ciliated cells. These cultures recapitulated the pseudostratified columnar epithelial architecture present in the native nasal epithelium and were permissive to respiratory virus infection. These results denote the potential of 3D bioprinting for high-throughput fabrication of nasal epithelial tissue models not only for infection studies but also for other purposes, such as disease modeling, immunological studies, and drug screening.