BV2 Cells - Central Nervous System Research Explained with BV2 Microglia Cells
BV2 is a mouse derived microglial cell line broadly used in neuroscience research. This immortalized cell line can serve as an in vitro model to study neurodegenerative diseases and associated cell conditions and processes, i.e., neuroinflammation. Besides, BV2 cells are considered an alternative model system for primary microglia.
This article will discuss the origin, cell culture information, and research applications of a mouse microglial cell line, BV2. Particularly, we will go through the following:
- Origin and general characteristics of the BV2 cells
- BV2 cell line: Culturing information
- Advantages & limitations of BV2 cells
- Applications of BV2 cell line in research
- BV2 cells: Research publications
- Resources for BV2 cell line: Protocols, Videos, and More
1. Origin and general characteristics of the BV2 cells
This section of the article explains BV2 cell line origin and the general features that distinguish it from other microglial cell lines. Herein, you will study: What are BV2 cells? Where are BV2 cells from? What is the size of a BV2 cell?
- BV2 microglial cell line was obtained from the microglia of neonatal (newborn) C57/BL6 The cell line was immortalized by infecting cells with J2 retrovirus carrying v-raf/v-myc oncogene [1].
- Unstimulated BV2 cells possess amoeba-like, hypertrophied morphology. This morphology exhibits a highly activated and inflammatory condition of BV2 cells compared to primary microglia [2].
- The diameter reported for the BV-2 cell line ranges between 10 to 15 μm.
BV2 Vs ECO 2 cell line
Both are mouse microglial cell lines but distinct from each other. The major difference is that BV2 was immortalized by genetic manipulation, whereas ECO 2 was spontaneously immortalized. Moreover, ECO 2 possesses the same general characteristics as BV2 but requires colony-stimulating factor-1 (CSF-1) supplementation for its culturing.
2. BV2 cell line: Culturing information
Before handling and maintaining a cell line culture, cell culturing information is crucial. This article section will help you know all the key points for culturing BV2 cell lines. Particularly, we will talk about the following: What is the BV2 doubling time? Which media is used to culture BV2 cells? Is the BV2 cell line adherent or suspension? How do you thaw BV2 cells?
Key Points for Culturing BV2 Cells
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Doubling Time: |
The BV2 microglial cells grow very fast, with an average BV2 doubling time of 34.5 hours. |
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Adherent or in Suspension: |
BV2 is a semi-adherent cell line. Cells exist both in suspension and adherent form. |
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Split Ratio: |
This semi-adherent microglial cell line is sub-cultured at a split ratio of 1:2 to 1:4. For passaging cells, suspended cells are collected in a culture tube. In contrast, adherent cells are washed with PBS and incubated with Accutase (dissociation solution). After 10 minutes, adherent and suspended cells are centrifuged together and harvested. These cells are then added into fresh growth media flasks according to the recommended split ratio. |
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Growth Medium: |
DMEM media is used to culture the BV2 cell line. BV2 DMEM is supplemented with 10% FBS, 1.5 g/L NaHCO3, 4.5 g/L Glucose, 4 mM L-Glutamine, and 1.0 mM Sodium pyruvate for ideal cell growth. Media is renewed 2 to 3 times per week. |
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Growth Conditions: |
BV2 cultures are maintained in a 37°C humidified incubator with a continuous 5% CO2 delivery. |
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Storage: |
Frozen BV2 cell vials are kept below -150 °C temperature either in the vapour phase of liquid nitrogen or in an electric freezer. |
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Freezing Process and Medium: |
CM-1 or CM-ACF freezing media are recommended for BV2 cell lines. Cells are frozen using a slow freezing process that permits only a 1°C temperature drop per minute to maintain cell viability. |
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Thawing Process: |
Frozen BV2 cell vial is rapidly agitated in a water bath (37°C) for 40 to 60 seconds until a small ice clump is left. Thawed cells are added with fresh growth media and centrifuged to eliminate freezing media components. Collected cells are again resuspended and poured into a culture flask for growth. |
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Biosafety Level: |
Biosafety level 1 is recommended for BV2 cell line culturing. |
3. Advantages & limitations of BV2 cells
Like other cell lines, BV2 cells are also associated with some advantages and limitations. Some of these are mentioned here.
Advantages
The advantages of the BV2 cell line include:
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Primary microglia-like features |
BV2 cells possess some primary microglia-like characteristics and are used as an alternative model to study microglial functions and responses. They express F4/80, CD11b, and Iba1, which are essential biomarkers of primary microglia. |
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Immortalization |
BV2 cells are immortalized, enabling them to undergo continuous growth. This characteristic makes them ideal for long-term cell culture experiments. |
Limitations
The limitations associated with BV2 cells are:
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Murine origin cell line |
The BV2 cell line is derived from mouse microglia. Research findings using BV2 cells may have limited applicability to human-specific diseases and research. |
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In vitro model |
BV2 cells serve as an in vitro model for studying microglial functions. However, it is important to note that they may not fully replicate the characteristics and complexity of microglial cells in the brain in vivo. |
4. Applications of BV2 cell line in research
BV2 cell line offers several applications in neuroscience research. Some common research uses of BV2 cells are mentioned in this section.
Neurodegenerative disease research: The murine microglial cell line, BV2 is a valuable research tool to study neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease, and multiple sclerosis. Researchers have studied neurotoxicity and disease pathology and evaluated therapeutic agents using BV2 cell lines. Such as, a study conducted in 2020 assessed the anti-inflammatory and neuroprotective effect of a natural hydroxystilbene, Rhaponticin, present in the Rheum rhaponticum plant, using lipopolysaccharide activated BV2 cells as Parkinson's disease model. The compound attenuates lipopolysaccharide (LPS) mediated BV2 activation by inhibiting nitric oxide synthase and reducing reactive oxygen species and proinflammatory mediators. Briefly, Rhaponticin exerts anti-inflammatory and neuroprotective effects on the LPS-induced microglial model (BV2) [3]. Similarly, a study investigated the involvement of signalling pathways in neuroinflammation. Researchers developed an inflammation model through lipopolysaccharide-mediated BV2 activation. They found that AKT/Nrf-2/HO-1-NF-κB signalling axis is involved in neuroinflammation. Besides, they also evaluated beta-naphthoflavone (BNF), a natural flavonoid, for its anti-inflammatory and neuroprotective effects by using this model. The compound exerted these therapeutic effects by inhibiting BV2 activation [4]. Likewise, the research used BV2 cells and studied the ameliorative effect of zonisamide drug on mitochondrial dysfunction in microglial cells. This study's findings support zonisamide's clinical use for Parkinson's disease treatment [5].
5. BV2 cells: Research publications
The following are some interesting and most cited research studies featuring BV2 cells.
This research is published in the Journal of Alzheimer's Disease (2015). The study proposed that mitochondrial damage-derived DAMP (damage-associated molecular pattern) mtDNA molecule can cause inflammatory changes in microglial cells (BV2). Thus, they may also contribute to Alzheimer's disease neuroinflammation.
This article published in FARMACIA (2021) used BV2 cells and determined the therapeutic effect of Huanglian Jiedu Decoction (HLJDD) on Alzheimer disease. The study found that HLJDD promotes BV2 amyloid-beta phagocytosis by upsurging Trm2 protein expression as validated through BV2 western blot analysis.
Alpha-synuclein activates BV2 microglia dependent on its aggregation state
This research article published in Biochemical and Biophysical Research Communications (2016) proposed that alpha-synuclein, a soluble protein in the adult central nervous system, can activate BV2 cells depending upon their aggregation state.
Exosomes of BV-2 cells induced by alpha-synuclein: important mediator of neurodegeneration in PD
This research was published in Neuroscience Letters in 2013. This study states that exosomes secreted from alpha-synuclein-activated BV2 microglial cells can be essential mediators of neurodegeneration in Parkinson's disease.
This study was published in Frontiers Cellular Neuroscience (2019). It proposed that Idebenone, an antioxidant, modulates microglial polarization and reduces inflammation in lipopolysaccharide-activated BV2 cells and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induced Parkinson's disease mice model.
6. Resources for BV2 cell line: Protocols, Videos, and More
The online resources available on BV2 are limited. Here are some of them.
- Subculturing of BV2 cell line: This website link consists of a brief protocol for subculturing of BV2 cell lines.
- Thawing frozen cells: This video helps you learn the basic protocol for thawing and culturing frozen cells.
The cell culture protocol for BV2 cells is mentioned here.
- BV2 cell culturing: This website link contains the BV2 cell culture protocol. In addition, it also provides cell culture media and freezing media compositions for the BV2 cell line.
References
- Wang, Y., Y. Peng, and H. Yan, Commentary: Neuroinflammatory In Vitro Cell Culture Models and the Potential Applications for Neurological Disorders. Front Pharmacol, 2021. 12: p. 792614.
- Sarkar, S., et al., Characterization and comparative analysis of a new mouse microglial cell model for studying neuroinflammatory mechanisms during neurotoxic insults. Neurotoxicology, 2018. 67: p. 129-140.
- Zhao, F., et al., Neuroprotective effect of rhaponticin against Parkinson disease: Insights from in vitro BV‐2 model and in vivo MPTP‐induced mice model. Journal of Biochemical and Molecular Toxicology, 2021. 35(1): p. e22631.
- Gao, X., et al., Beta-naphthoflavone inhibits LPS-induced inflammation in BV-2 cells via AKT/Nrf-2/HO-1-NF-κB signaling axis. Immunobiology, 2020. 225(4): p. 151965.
- Tada, S., et al., Zonisamide Ameliorates Microglial Mitochondriopathy in Parkinson's Disease Models. Brain Sciences, 2022. 12(2): p. 268.