Florent Foisset, Christine Lehalle, Amel Nasri, Carine Bourdais, Cécilia Urena, Lisa Morichon, Agathe Cœur, Aurélie Petit, Isabelle Vachier, Quentin Muller, Vincent Flacher, Jamila Chakir, Gladys Massiera, Vimala Sridurai, Engi Ahmed, Said Assou, Arnaud Bourdin, Nelly Frossard, John De Vos
Primary human bronchial fibroblasts seeded into collagen-chitosan matrix dramatically improved iALI epithelial differentiation from vAFE cells, related to highly specific transcriptomic signatures when compared to iPSC derived fibroblasts.
RATIONALE: Derived iPSCs airway epithelium are challenging given their dependency on the mesenchymal compartment. We hypothesized that growing vAFE cells on well-organized stiff matrix precolonized by adult pulmonary fibroblasts would improve epithelial differentiation yield and maturity.
METHODS: Collagen-1/chitosan matrix were engineered to reach stiffness and scaffolding characteristics of subepithelial compartments. Primary fibroblasts derived from human lung samples were seeded for 45 days before the addition of vAFE cells differentiated from iPSCs, and comparisons made with iPSC-derived fibroblasts. Beads tracking was used to assess cilia beating efficiency.
RESULTS: Primary human bronchial fibroblasts were able to enrich the CC Matrix with extracellular matrix components such as collagen, decorin and vimentin. In turn, iALI cultures performed in primary fibroblasts enriched CC matrix successfully led to high level of epithelial differentiation including rare cells (club, basal, neuroendocrine, ciliated, secretory). Large apical surfaces were covered by approximately 60% of ciliated cells able to generate mucociliary vortex.
CONCLUSION: Primary human bronchial fibroblasts seeded into collagen-chitosan matrix dramatically improved iALI epithelial differentiation from vAFE cells, related to highly specific transcriptomic signatures when compared to iPSC derived fibroblasts.