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The HT22 cell line, an immortalized subclone derived from HT4 cells of the mouse hippocampus, is pivotal in neuropharmacological research. Originating through the immortalization of mouse neuronal tissues with a temperature-sensitive SV40 T-antigen, HT22 cells offer a unique in vitro model to investigate the mechanisms underlying glutamate-induced cytotoxicity, which plays a significant role in neurodegenerative disorders such as Alzheimer's, Huntington's, and Parkinson's diseases.
HT22 cells exhibit a neuronal phenotype and are highly sensitive to glutamate, an essential excitatory neurotransmitter involved in critical brain functions like cognition, learning, and memory. However, excessive glutamate intake can lead to glutamate toxicity and overexcitation of nerve cells, causing cell damage or death through mechanisms that involve oxidative stress and apoptosis.
HT22 mouse hippocampal cells are employed in neurotoxicity studies, such as those examining the effects of isoflurane exposure, for exploring the chromatin landscape and epigenetic signatures, and to examine the effects of serotonergic input on hippocampal neurogenesis. The latter includes the study of serotonin reuptake inhibitors and their role in antidepressant screening, as well as the impact of serotonin transporter (SERT) glycosylation on neuronal function.
The HT22 cell line, with its well-characterized response to glutamate and its utility in studying the serotonergic system, continues to be a valuable tool in the advancement of neuropharmacology and the development of treatments for a range of neurological disorders.
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- cultureMedium: DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- freezeMedium: As a cryopreservation medium, use 50% basal medium + 40% FBS + 10% DMSO, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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