Mammalian Reoviruses: Propagation, Quantification, and Storage

Abstract

Mammalian reoviruses are pathogens that cause gastrointestinal and respiratory infections. In humans, the mammalian reoviruses usually cause mild or subclinical disease, and they are ubiquitous, with most people mounting immunity at a young age. Reoviruses are prototypic representations of the Reoviridae family, which contains many highly pathogenic viruses. This article describes techniques for culturing mouse fibroblast L929 cell lines, the preferred cell line in which most mammalian reovirus studies take place. In addition, mammalian reovirus propagation, quantification, purification, and storage are described. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Propagation of mammalian reoviruses in cell culture from virus stocks. Alternate Protocol 1: Large-scale propagation (and purification) of mammalian reoviruses in cell culture from virus stocks. Basic Protocol 2: Quantification of mammalian reoviruses by plaque assay with neutral red staining. Alternate Protocol 2: Quantification of mammalian reoviruses by plaque assay with crystal violet staining. Basic Protocol 3: Storage of mammalian reoviruses. Support Protocol 1: Growth and maintenance of mouse L929 cells. Support Protocol 2: Plating L929 cells.