Abstract
Microglia are macrophage-like cells that populate the central nervous system (CNS) and become activated upon injury or infection. Microglia have been implicated as playing critical roles in various CNS diseases including multiple sclerosis (MS), a human autoimmune demyelinating disease, as well as in other neurodegenerative diseases. Two well-characterized models of MS, relapsing experimental autoimmune encephalomyelitis (R-EAE) and Theiler's murine encephalomyelitis virus (TMEV)-induced demyelinating disease, are inducible in SJL mice and model the relapsing-remitting and chronic-progressive forms of MS, respectively. These models are useful for the study of the mechanisms of initiation, progression, and therapy of the disease. Currently, a major limitation to studying the functions of microglia in these murine models of MS is the restricted number of cells capable of being isolated from the CNS of neonatal mice and propagated in culture. The current studies describe the preparation of SV-40 large T antigen-immortalized mouse microglia lines, M4T.4 and M4T.6, from the SJL/J mice. The immortalization technique was very efficient requiring only 6 weeks to develop long-term, highly replicating cell lines. The resulting microglia cell lines remain quiescent, but are induced to express various immune cytokines and to function as efficient antigen presenting cells upon activation with IFN-γ or infection with TMEV. Thus, the SV-40 large T antigen immortalized microglia lines react to innate and infectious stimuli similar to primary microglia isolated from neonatal mice, but are more easily maintained in culture. This technique should allow for the efficient cultivation of large numbers of microglial cells from a variety of disease-relevant mouse strains, including knock-out and transgenic mice.
Original language | English (US) |
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Pages (from-to) | 33-43 |
Number of pages | 11 |
Journal | Journal of Neuroscience Methods |
Volume | 128 |
Issue number | 1-2 |
DOIs | |
State | Published - Sep 30 2003 |
Bibliographical note
Funding Information:This work was supported in part by USPHS NIH Grants NS-23349 and NS-30871. J.K. Olson is supported by an Advanced Postdoctoral Fellowship Grant FA 1517 from the National Multiple Sclerosis Society. The authors thank Marissa McShane and Wendy Smith Begolka for their technical assistance.
Keywords
- Antigen presentation
- Immortalization
- Microglial cell line
- Theiler's virus