Abstract
The innate immune system is evolutionarily conserved and shared by a wide spectrum of cells, including epithelial cells, endothelial cells, fibroblasts, macrophages, dendritic cells, and lymphocytes. The extensive utilization of genetically manipulated animals in innate immunity studies has become the standard approach to confirm functional data acquired in cell lines. The easy generation and physiological relevance of mouse embryonic fibroblasts (MEFs) have made them a powerful tool in discovering novel signaling pathways, investigating regulatory networks, and exploring biochemical profiling of protein complexes involved in innate immune responses. Due to their extensive use, this chapter aims to provide a protocol for generating, maintaining, and storing primary MEFs for those who have minimal experience in animal models. Precautions and notes are integrated into the description of each step of the protocol for the benefits of minimizing unnecessary cross-referencing.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Davis BK, Wen H, Ting JP (2011) The inflammasome NLRs in immunity, inflammation, and associated diseases. Annu Rev Immunol 29:707–735
Ting JP, Duncan JA, Lei Y (2010) How the noninflammasome NLRs function in the innate immune system. Science 327:286–290
Wen H, Lei Y, Eun SY, Ting JP (2010) Plexin-A4-semaphorin 3A signaling is required for Toll-like receptor- and sepsis-induced cytokine storm. J Exp Med 207:2943–2957
Ishikawa H, Barber GN (2008) STING is an endoplasmic reticulum adaptor that facilitates innate immune signalling. Nature 455:674–678
Seth RB, Sun L, Ea CK, Chen ZJ (2005) Identification and characterization of MAVS, a mitochondrial antiviral signaling protein that activates NF-kappaB and IRF 3. Cell 122:669–682
Yoneyama M, Kikuchi M, Natsukawa T, Shinobu N, Imaizumi T et al (2004) The RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses. Nat Immunol 5:730–737
Kawai T, Takahashi K, Sato S, Coban C, Kumar H et al (2005) IPS-1, an adaptor triggering RIG-I- and Mda5-mediated type I interferon induction. Nat Immunol 6:981–988
Meylan E, Curran J, Hofmann K, Moradpour D, Binder M et al (2005) Cardif is an adaptor protein in the RIG-I antiviral pathway and is targeted by hepatitis C virus. Nature 437:1167–1172
Allen IC, Moore CB, Schneider M, Lei Y, Davis BK et al (2011) NLRX1 protein attenuates inflammatory responses to infection by interfering with the RIG-I-MAVS and TRAF6-NF-kappaB signaling pathways. Immunity 34:854–865
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Lei, Y. (2013). Generation and Culture of Mouse Embryonic Fibroblasts. In: Allen, I. (eds) Mouse Models of Innate Immunity. Methods in Molecular Biology, vol 1031. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-481-4_7
Download citation
DOI: https://doi.org/10.1007/978-1-62703-481-4_7
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-480-7
Online ISBN: 978-1-62703-481-4
eBook Packages: Springer Protocols