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Updated: Jul 2, 2026
Magnetic Isolation of Microglial Cells from Neonate Mouse for Primary Cell Cultures
Published on: July 25, 2022
Differential migration, LPS-induced cytokine, chemokine, and NO expression in immortalized BV-2 and HAPI cell lines
Ryan J Horvath1, Nancy Nutile-McMenemy, Matthew S Alkaitis
1Department of Pharmacology, Dartmouth Medical School, Hanover, New Hampshire, USA.
Abstract:
Microglial cells are hematopoietically derived monocytes of the CNS and serve important neuromodulatory, neurotrophic, and neuroimmune roles. Following insult to the CNS, microglia develop a reactive phenotype, migrate to the site of injury, proliferate, and release a range of proinflammatory, anti-inflammatory, and neurotrophic factors. Isolation of primary microglial cell cultures has been an integral step in elucidating the many roles of these cells. In addition to primary microglial cells, several immortalized cell lines have been created to model primary microglia in vitro, including murine-derived BV-2 cells and rat derived highly aggressive proliferating immortalized (HAPI) cells. Here, we compare rat primary microglial, BV-2, and HAPI cells in experiments assessing migration, expression of activation markers, and production and release of nitric oxide, cytokines, and chemokines. BV-2 and HAPI cells responded similarly to primary microglia in experiments assessing migration, ionized calcium binding adaptor molecule 1 expression, and nitric oxide release. However, BV-2 and HAPI cells did not model primary microglia in experiments assessing tumor necrosis factor-alpha, interleukin-1beta, interleukin-6, and monocyte chemoattractant protein-1 expression and release and phospho-extracellular signal-regulated kinase 44/42 expression following lipopolysaccharide treatment. These results indicate that BV-2 and HAPI cell cultures only partially model primary microglia and that their use should therefore be carefully considered.
Insights
Researchers compared primary microglia with BV-2 and HAPI cell lines. While these cell lines partially mimic primary microglia responses, they do not fully replicate key inflammatory markers, suggesting cautious use in CNS research.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia are key immune cells in the central nervous system (CNS) with critical roles in neuromodulation, neurotrophic support, and neuroimmunity.
- Upon CNS injury, microglia exhibit reactive phenotypes, migrating to injury sites, proliferating, and releasing various signaling molecules.
- Primary microglial cultures are essential for studying these cells, but immortalized cell lines like BV-2 (murine) and HAPI (rat) are also used as models.
Purpose of the Study:
- To compare the utility of rat primary microglial cells with two commonly used immortalized cell lines, BV-2 and HAPI.
- To assess the similarities and differences in migration, activation marker expression, and the release of nitric oxide, cytokines, and chemokines between primary microglia and cell lines.
Main Methods:
- Primary rat microglia, BV-2, and HAPI cells were cultured.
- Experiments assessed cell migration, expression of ionized calcium binding adaptor molecule 1 (Iba1), nitric oxide (NO) production, and the release of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β), interleukin-6 (IL-6), and monocyte chemoattractant protein-1 (MCP-1) after lipopolysaccharide (LPS) stimulation.
- Phospho-extracellular signal-regulated kinase 44/42 (p-ERK44/42) expression was also evaluated.
Main Results:
- BV-2 and HAPI cells showed similar responses to primary microglia in migration assays, Iba1 expression, and NO release.
- However, significant discrepancies were observed in the expression and release of TNF-α, IL-1β, IL-6, and MCP-1 following LPS treatment.
- p-ERK44/42 expression also differed between primary microglia and the cell lines.
Conclusions:
- BV-2 and HAPI cell lines only partially replicate the behavior of primary microglia.
- These immortalized cell lines may not be suitable models for all aspects of microglial activation, particularly inflammatory responses.
- Researchers should carefully consider the limitations of BV-2 and HAPI cells when designing experiments investigating microglial function in the CNS.
