ArticleVirology journal2014
Resolving the titer of murine cytomegalovirus by plaque assay using the M2-10B4 cell line and a low viscosity overlay.
Article in Virology journal, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 44 papers.
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Who cites it
44 citing papers in PubMed, 53 citations in OpenAlex.
- Age determines NK cell fate and tissue compartmentalization to CMV infection.bioRxiv : the preprint server for biology · 2026Article
- Persistent microglial activation following neonatal CMV infection mediates neurodegeneration.Science advances · 2026Article
- The impact of cytomegalovirus coinfection on tuberculosis in the mouse model.BMC infectious diseases · 2026Article
- Age-driven shifts in T and NK cell responses amplify inflammation and coagulopathy during viral infection in mice and humans.Frontiers in immunology · 2026Article
- Leukotriene B4 regulates T cell recognition and control of MCMV in mucosal tissues.Mucosal immunology · 2025Article
- Immune responses drive chorioretinitis and retinal pathology after neonatal CMV infection.Science advances · 2024Article
- Intermittent cytomegalovirus infection alters neurobiological metabolism and induces cognitive deficits in mice.Brain, behavior, and immunity · 2024Article
- Systemic immunostimulation induces glucocorticoid-mediated thymic involution succeeded by rebound hyperplasia which is impaired in aged recipients.Frontiers in immunology · 2024Article
- Novel Mouse Model of Murine Cytomegalovirus-Induced Adaptive NK Cells.ImmunoHorizons · 2022Article
- Article
- Mechanisms of Antiviral Cytotoxic CD4 T Cell Differentiation.Journal of virology · 2021Article
- The degree of polymerization and sulfation patterns in heparan sulfate are critical determinants of cytomegalovirus entry into host cells.PLoS pathogens · 2021Article
- NUDT15 polymorphism influences the metabolism and therapeutic effects of acyclovir and ganciclovir.Nature communications · 2021Article
- Influenza- and MCMV-induced memory CD8 T cells control respiratory vaccinia virus infection despite residence in distinct anatomical niches.Mucosal immunology · 2021Article
- Aberrant type 1 immunity drives susceptibility to mucosal fungal infections.Science (New York, N.Y.) · 2021Article
- Inhibitory Molecules PD-1, CD73 and CD39 Are Expressed by CD8Frontiers in immunology · 2021Article
- Hematopoietic cell-mediated dissemination of murine cytomegalovirus is regulated by NK cells and immune evasion.PLoS pathogens · 2021Article
- A Review of Murine Cytomegalovirus as a Model for Human Cytomegalovirus Disease-Do Mice Lie?International journal of molecular sciences · 2020Review
- Platelet-endothelial associations may promote cytomegalovirus replication in the salivary gland in mice.Platelets · 2020Article
- Chronic viral infections impinge on naive bystander CD8 T cells.Immunity, inflammation and disease · 2020Article
Corrections and comments
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Authors and funding
3 authors at 2 institutions in 1 country.
Funding
Abstract
backgroundMurine cytomegalovirus (MCMV) is increasingly used as an infectious model to investigate host-pathogen interactions in mice. Detailed methods have been published for using primary murine embryonic fibroblasts (MEFs) for preparing stocks and determining viral titers of MCMV. For determining the titer of MCMV by plaque assay, these methods rely on a high viscosity media that restricts viral spreading through the supernatant of the culture, but is also usually too viscous to pipet. Moreover, MEFs must be repeatedly generated and can vary widely from batch-to-batch in purity, proliferation rates, and the development of senescence. In contrast, the M2-10B4 bone marrow stromal cell line (ATCC # CRL-1972), which is also permissive for MCMV, has been reported to produce high-titer stocks of MCMV and has the considerable advantages of growing rapidly and consistently. However, detailed methods using these cells have not been published.
methodsWe modified existing protocols to use M2-10B4 cells for measuring MCMV titers by plaque assay.
resultsWe found that MCMV plaques could be easily resolved on monolayers of M2-10B4 cells. Moreover, plaques formed normally even when cultures of M2-10B4 cells were less than 50% confluent on the day of infection, as long as we also used a reduced viscosity overlay.
conclusionsOverall, our protocol enabled us to use a consistent cell line to assess viral titers, rather than repeatedly producing primary MEFs. It also allowed us to start the assay with 4-fold fewer cells than would be required to generate a confluent monolayer, reducing the lead-time prior to the start of the assay. Finally, the reduced viscosity CMC could be handled by pipet and did not need to be pre-mixed with media, thus increasing its shelf-life and ease-of-use. We describe our results here, along with detailed protocols for the use of the M2-10B4 cell lines to determine the titer and grow stocks of MCMV.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the Socratic graph.