Creation of a spatially complex mucus bilayer on an in vitro colon model

• Published in: Scientific reports Vol. 14; no. 1; pp. 16849 - 15
• Main Authors: Villegas-Novoa, Cecilia, Wang, Yuli, Sims, Christopher E, Allbritton, Nancy L
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 22.07.2024; Nature Publishing Group UK; Nature Portfolio
• Subjects: Air–liquid interface; Bacteria; Cell culture; Cells, Cultured; Colon; Colon - metabolism; Colon-on-chip; Crypt; Epithelium; Goblet cell; Goblet cells; Goblet Cells - metabolism; Humans; Hydrogels; Inflammatory bowel disease; Intestinal Mucosa - metabolism; Lectins; Microphysiologic system; Models, Biological; Mucus; Mucus - metabolism; Pattern formation; Physiology; Population studies; Regions; Secretion; Stem cells; Goblet cell; Mucus; Microphysiologic system; Crypt; Air–liquid interface; Colon-on-chip
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-024-67591-9

The colonic epithelium is comprised of three-dimensional crypts (3D) lined with mucus secreted by a heterogeneous population of goblet cells. In this study, we report the formation of a long-lived, and self-renewing replica of human 3D crypts with a mucus layer patterned in the X-Y-Z dimensions. Primary colon cells were cultured on a shaped scaffold under an air-liquid interface to yield architecturally accurate crypts with a mucus bilayer (605 ± 180 μm thick) possessing an inner (149 ± 50 μm) and outer (435 ± 111 μm) region. Lectins with distinct carbohydrate-binding preferences demonstrated that the mucus in the intercrypt regions was chemically distinct from that above and within the crypts replicating in vivo chemical patterning. Constitutive mucus secretion ejected beads from crypt lumens in 8-10 days, while agonist-stimulated secretion increased mucus thickness by 17-fold in 8 h. The tissue was long-lived, > 50 days, the longest time assessed. In conclusion, the in vitro mucus replicated key physiology of the human mucus, including the bilayer (Z) structure and intercrypt-crypt (X-Y) zones, constitutive mucus flow, spatially complex chemical attributes, and mucus secretion response to stimulation, with the potential to reveal local and global determinants of mucus function and its breakdown in disease.