The human microglia (HMC-3) as a cellular model of neuroinflammation

Izabela Lepiarz-Raba, Olumayokun Olajide

Research output: Contribution to journalMeeting Abstractpeer-review


Introduction: Neuroinflammation has been shown not only as a crucial component but the cause of Alzheimer disease and other neurodegenerative disorders. Microglia are primary immunocompetent cells in the brain, therefore they are widely used to study neuroinflammation and test anti-inflammatory properties of compounds. Current studies of microglial activation are widely performed with rodent models. However, development of human microglial cell model will overcome species-specific differences and will bring us considerably closer to development of effective investigation of neuroinflammation.

Method: HMC-3 cells were stimulated with IFNγ (10 ng/ml) or TNFα (25 ng/ml) at different time points. The phosphorylation levels of NF-κB p65, ERK1/2 and p38 were assessed using western blots. Additionally immunofluorescence staining of NF-κB p65 subunit was performed. In order to confirm NF-κB activation cells were pre-incubated with BAY 11-7082 for 30 min and subsequently stimulated with IFNγ. The production of IL-6 was measured using ELISA and level of TLR4, TLR9 and Iba1 expression was assessed using immunofluorescence staining.

Results: Stimulation of HMC-3 cells with IFNγ caused phosphorylation of NF-κB p65 and pre-incubation with BAY 11-7082 reduced its level. IFNγ also triggered phosphorylation of ERK1/2 and p38. Similarly, pre-incubation of HMC-3 cells with TNFα increased phosphorylation of NF-κB p65, ERK1/2 and p-38. IFNγ caused 1.9-fold upregulation of IL-6, whereas TNFα increased IL-6 production more than 6-fold. IFNγ and TNFα enhanced expression of Iba1. HMC-3 cells express higher level of TLR9 than TLR4.

Conclusions: IFNγ and TNFα induce activation of HMC-3 triggering NF-κB, ERK1/2 and p38 pathways. Activation of HMC-3 cells with TNFα leads to greater inflammatory response than IFNγ stimulation. HMC-3 cell line is suitable to examine anti-inflammatory properties and mechanism of action of anti-inflammatory agents. Further experiments will be carried out to investigate the role of toll-like receptors (TLRs) in inflammatory signalling cascades in the human microglia.
Original languageEnglish
Pages (from-to)S92
Number of pages1
JournalIBRO Reports
Issue numberSupplement
Early online date20 Sep 2019
Publication statusPublished - 20 Sep 2019
EventThe 10th IBRO World Congress of Neuroscience: Joint Meeting of International Brain Research Organization & Federation of Asian-Oceanian Neuroscience Societies - Daegu, Korea, Republic of
Duration: 21 Sep 201925 Sep 2019
Conference number: 10


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