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Published on: June 14, 2020
Characterization and comparative analysis of a new mouse microglial cell model for studying neuroinflammatory
Souvarish Sarkar1, Emir Malovic1, Deeksha Sarda1
1Parkinson Disorders Research Laboratory, Iowa Center for Advanced Neurotoxicology, Department of Biomedical Sciences, 2062 Veterinary Medicine Building, Iowa State University, Ames, IA, 50011, United States.
Abstract:
Microglia are the first responders of the central nervous system, acting as the key modulators of neuroinflammation observed during neurotoxic insults as well as in the pathophysiology of several neurodegenerative disorders including Alzheimer's (AD), Parkinson's (PD), and Huntington's diseases (HD). The number of publications on microglia has increased steadily throughout the past decade because of immense interests in the neuroinflammation that precedes the neurodegenerative process. To study microglial biology and its role in modulating neuroinflammation, immortalized microglial cell lines derived from mice, rats, and humans have been developed. Among these, the BV2 mouse microglial cell line is the most well characterized and widely used cell culture model. However, even unstimulated BV2 cells exhibit an amoeboid, hypertrophied morphology, indicating a highly activated and inflammatory state compared to primary microglia, thus making them less than ideal for studying the low-dose effects of toxicants on microglial activation. Therefore, we performed an in-depth characterization of a recently developed mouse microglial cell (MMC) line, which we compared with primary mouse microglia (PMG) and BV2s to identify which cell line was best suited for studying the microglial response to neurotoxicants. Comparative analyses reveal that MMCs are strikingly more similar to PMGs in basal activity, morphology, and sensitivity, than are BV2s. Furthermore, basal nitrite and inflammatory cytokine levels are significantly higher in BV2s compared to MMCs. BV2 cells are also less reactive to the inflammagen LPS compared to MMCs, due to the higher basal activation state of BV2s. Collectively, our in-depth analyses of morphology, basal activity, and responsivity to two different stimuli (LPS, aggregated α-synuclein) demonstrate that MMCs closely mimic neonatal PMGs, and are discernibly more suitable than BV2s for studying the neuroinflammatory mechanisms of neurotoxicants.
Insights
A new mouse microglial cell (MMC) line closely mimics primary microglia, offering a superior model for studying neuroinflammation and neurotoxicants compared to the widely used BV2 cell line.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia are key modulators of neuroinflammation in neurodegenerative diseases like Alzheimer's and Parkinson's.
- Immortalized microglial cell lines are used to study microglial biology.
- The BV2 cell line, while common, exhibits high basal activation, limiting its use for studying low-dose toxicant effects.
Purpose of the Study:
- To characterize a new mouse microglial cell (MMC) line.
- To compare the MMC line with primary mouse microglia (PMG) and BV2 cells.
- To determine the most suitable cell line for studying microglial responses to neurotoxicants.
Main Methods:
- In-depth characterization of MMC, PMG, and BV2 cell lines.
- Comparative analysis of morphology, basal activity, and inflammatory marker levels (nitrite, cytokines).
- Assessment of cellular responsiveness to stimuli like lipopolysaccharide (LPS) and aggregated alpha-synuclein.
Main Results:
- MMCs exhibit basal activity, morphology, and sensitivity more similar to PMGs than BV2s.
- BV2 cells show significantly higher basal nitrite and inflammatory cytokine levels than MMCs.
- BV2 cells display reduced reactivity to LPS compared to MMCs due to their higher basal activation.
Conclusions:
- The MMC line closely mimics neonatal primary mouse microglia.
- MMCs are more suitable than BV2 cells for investigating neuroinflammatory mechanisms of neurotoxicants.
- This finding aids in selecting appropriate cell models for neuroinflammation research.
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