Palor, Machaela;
Haughey, Elizabeth K;
Herbert, Jenny;
O'Callaghan, Christopher;
Smyth, Rosalind L;
De Coppi, Paolo;
Hynds, Robert E;
(2025)
Ciliated Epithelial Cell Differentiation at Air-Liquid Interface and RSV infection using Animal-Free Media and Substrates.
ERJ Open Research
00028-2025.
10.1183/23120541.00028-2025.
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Abstract
Animal-derived components in cell culture, such as foetal bovine serum (FBS) and extracellular matrix proteins (ECM), pose ethical concerns and contribute to variability in experimental outcomes. This study explores the use of animal-free cell culture media and substrates to support the growth and differentiation of primary human bronchial epithelial cells (BECs), as well as their infection by respiratory syncytial virus (RSV). We evaluated the performance of jellyfish collagen 0 and recombinant ECM proteins as alternatives to traditional mammalian substrates. Additionally, we assessed the use of animal-free media and human serum (HS) in viral propagation using HEp2 cells. Results demonstrate that the use of animal-free medium and matrix proteins and human serum can support primary epithelial cell growth and differentiation, with high-levels of ciliation and barrier integrity. RSV propagation in animal-free medium produced an increase in viral titres, indicating the potential of these systems for anti-viral research. Transitioning to include more animal-free medium and substrates for primary cell culture and viral propagation will help improve the ethical standing of research and offer more human-relevant models for studying viral diseases in the future.
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