Differential migration, LPS-induced cytokine, chemokine, and NO expression in immortalized BV-2 and HAPI cell lines and primary microglial cultures.
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.
Horvath
Ryan J
RJ
Department of Pharmacology, Dartmouth Medical School, Hanover, New Hampshire, USA.
Nutile-McMenemy
Nancy
N
Alkaitis
Matthew S
MS
Deleo
Joyce A
JA
D000818:Animals; D000831:Animals, Newborn; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects*; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D016207:Cytokines / Q000235:genetics / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology*; D051379:Mice; D008840:Microfilament Proteins; D017628:Microglia / Q000187:drug effects* / Q000502:physiology*; D008911:Minocycline / Q000494:pharmacology; D018696:Neuroprotective Agents / Q000494:pharmacology; D009569:Nitric Oxide / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D013997:Time Factors; D014970:Xanthines / Q000494:pharmacology
C100745:Aif1 protein, rat; D002135:Calcium-Binding Proteins; D016207:Cytokines; D008070:Lipopolysaccharides; D008840:Microfilament Proteins; D018696:Neuroprotective Agents; D014970:Xanthines; D009569:Nitric Oxide; C032114:propentofylline; D048049:Extracellular Signal-Regulated MAP Kinases; D008911:Minocycline