Ciliated epithelial cell differentiation at air-liquid interface and respiratory syncytial virus infection using animal-free media and substrates

利用无动物源培养基和底物,在气液界面诱导纤毛上皮细胞分化和呼吸道合胞病毒感染

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作者:Machaela Palor ,Elizabeth K Haughey ,Jenny Herbert ,Christopher O'Callaghan ,Rosalind L Smyth ,Paolo De Coppi ,Robert E Hynds ,Claire M Smith

Abstract

Background: Animal-derived components in cell culture, such as fetal bovine serum and extracellular matrix proteins, 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, as well as their infection by respiratory syncytial virus (RSV). Methods: We evaluated the performance of jellyfish collagen 0 and recombinant extracellular matrix proteins as alternatives to traditional mammalian substrates. Additionally, we assessed the use of animal-free medium and human serum (HS) in viral propagation using HEp2 cells. Results: The use of animal-free medium, matrix proteins and HS 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 antiviral research. Conclusion: 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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