Mammalian Reoviruses: Propagation, Quantification, and Storage

Abstract

AbstractMammalian 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 stocksAlternate Protocol 1: Large‐scale propagation (and purification) of mammalian reoviruses in cell culture from virus stocksBasic Protocol 2: Quantification of mammalian reoviruses by plaque assay with neutral red stainingAlternate Protocol 2: Quantification of mammalian reoviruses by plaque assay with crystal violet stainingBasic Protocol 3: Storage of mammalian reovirusesSupport Protocol 1: Growth and maintenance of mouse L929 cellsSupport Protocol 2: Plating L929 cells