HT22 cells were purchased from ATCC (Manassas, VA, USA). Cells were transferred to a DMEM medium containing 10% fetal bovine serum and 1% penicillin/streptomycin at 37 °C with 5% CO2. Upon confluent of 70%-80%, the cells were synchronized with serum-free SFM medium for 24 h and then treated with or without oxygen and glucose deprivation (OGD) for 1 h.
Ht22 cells
Manufactured by American Type Culture Collection
Sourced in United States
The HT22 cell line is a mouse hippocampal neuronal cell line commonly used in neuroscience research. The cells are derived from the hippocampus of a mouse and are immortalized, allowing for long-term culture and study. The HT22 cells maintain several characteristics of hippocampal neurons and are a valuable tool for investigating various aspects of neuronal function and biology.
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Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Pc12 cells, by American Type Culture Collection (2 mentions)
Sodium glutamate, by Merck Group (1 mentions)
Lc3a b, by Abcam (1 mentions)
P drp1, by Abcam (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Sodium selenite, by Merck Group (1 mentions)
β actin, by Abcam (1 mentions)
15nh4cl, by Cambridge Isotopes (1 mentions)
Rabbit anti gfp antibody, by Cell Signaling Technology (1 mentions)
Bicinchoninic acid bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Rabbit anti na k atpase antibody, by Cell Signaling Technology (1 mentions)
Phorbol 12 13 dibutyrate pdbu, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by ZenBio (1 mentions)
Rabbit anti β actin, by Cell Signaling Technology (1 mentions)
Glutathione sepharose 4b, by GE Healthcare (1 mentions)
Phorbol 12 myristate 13 acetate, by LC Laboratories (1 mentions)
1 2 dioctanoyl sn glycerol, by Avanti Polar Lipids (1 mentions)
Bradford protein assay, by Bio-Rad (1 mentions)
U87mg cells, by American Type Culture Collection (1 mentions)
16 protocols using ht22 cells
1
Evaluating Cellular Responses to OGD
2
Mitigating VPA-induced Toxicity in HT-22 Cells
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HT-22 cells (immortalized hippocampal neuron cells), a surrogate of neuroepithelial cells, were obtained from ATCC (Manassas, USA), and cultured in DMEM containing 10% FBS. To explore the protective effect of MT on the toxic effects of VPA, HT-22 cells were cotreated with 10 mM VPA and 20 μM MT for 48 h. Cultured cells were harvested for further analysis.
3
Alzheimer's Disease In Vitro Model
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Cell cultures were generated using immortalised mouse hippocampus HT-22 cells, purchased from the ATCC (Manassas, VA, USA). HT-22 was cultured in Dulbecco's Modified Eagle medium supplemented with 10% (v/v) heat-inactivated foetal bovine serum, 25 mM glucose, 2 mM l-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C in a humidified atmosphere containing 95% (v/v) air and 5% (v/v) CO2. The cells were seeded at a density of 2.0 × 105 cell/well in 6-well plates. To induce the Alzheimer’s in vitro model, 20 μM Aβ42 was treated at 24 h in the serum-free condition. And then HT-22 cells were treated with 3 days of rMS stimulation for 20 min daily. The control group was incubated for 72 h.
4
Murine Hippocampal Neuronal HT22 Cell Culture
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Murine hippocampal neuronal HT22 cells (originally purchased from ATCC) were cultured in Dulbecco’s Modified Eagle Medium (DMEM)/F12 (Invitrogen, Carlsbad, CA), supplemented with 10% fetal bovine serum (FBS, HyClone), 100 nM penicillin/streptomycin, at 37 °C in 5% CO2 and 90% relative humidity. The culture medium was renewed every 2 days. Sodium glutamate, and sodium selenite (Sigma-Aldrich, St Lois, MO), were reconstituted in water and diluted to appropriate concentrations in cell culture medium. Antibodies directed against caspase-3, caspase-9, apoptosis protease-activating factor-1 (Apaf-1), cleaved caspase-3, cleaved caspase-9 were purchased from Cell Signaling Technology, Danvers, MA. Anti LC3A/B, Fis1, Drp1, p-Drp1, and β-actin were purchased from Abcam, Cambridge, MA.
5
Lipid and Protein Reagents for Cell Signaling Studies
6
Modeling Cerebral Ischemia/Reperfusion Injury
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The mouse hippocampal neuronal HT22 cells were purchased from ATCC. They were cultured in a complete medium (high glucose DMEM medium, 10% fetal bovine serum, and 1% penicillin and streptomycin). To mimic cerebral ischemia/reperfusion injury in vitro, HT22 cells were exposed to oxygen-glucose deprivation for 9 h and reoxygenation (OGD/R) according to a previous protocol (Jiang et al., 2000 (link)). For OGD/R, the medium was changed to glucose-free DMEM supplemented with 10% fetal bovine serum and 1% penicillin and streptomycin and cultured in an incubator perfused with O2/CO2 (1.2%/5%) tri-gas. After 9 h of OGD, the cells were replaced with the complete medium and cultured in the incubator perfused with 21% O2/5%CO2 for 24 h to induce reoxygenation. These cell experiments were also divided into three groups: control, oxygen-glucose deprivation/reoxygenation (labeled as OGD), and OGD plus 10 μg/ml MC-ELNs (labeled as MC-ELNs, treated before and after 9 h of OGD).
7
Assessing CYBA's Cytotoxicity in Neuronal and Glioblastoma Cells
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The HT22 cells (mouse hippocampal neuronal cell line, ATCC, Manassas, United States) and U87MG cells (human malignant glioblastoma cells, ATCC, Manassas, United States) were cultured in Dulbecco’s modified Eagle’s medium (DMEM), which was supplemented with 10% FBS (WelGene, Daegu, Korea). The cells were incubated at a temperature of 37°C under a humidified atmosphere containing 5% CO2. The cells were subsequently cultivated on 96-well culture plates, with a density of 1 × 104 cells per well. After 24 h of incubation, cells were treated with various doses of CYBA fo r 24 h before being cultured with 10% CCK-8 solution for 2 h. Afterwards, the optical density values were assessed at a wavelength of 450 nm.
8
Bergenin Protects Neuronal Cells
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The neuronal HT-22 cells and PC-12 cells were obtained from American Type Culture Collection (ATCC) (Manassas, VA). HT-22 and PC-12 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS). These cells were maintained in the presence of 100 µg/mL streptomycin and 100 U/mL penicillin at 37 °C in a humidified atmosphere containing 5% CO2 and 95% air. Stock solution of 100 mM was prepared by dissolving the sample (Bergenin) in dimethyl sulfoxide (DMSO). Which was diluted further for working concentrations, i.e., 1, 10, 50 and 100 µM of Bergenin, keeping the final concentration of DMSO was < 0.2% to avoid any interference with the assay. Upon achieving the cell density of 70–80%, HT-22 and PC-12 cells were exposed to the indicated concentration of Bergenin for 2 h. The cells were then treated with H2O2 for 24 h. Then various assays were performed.
9
Oxygen-Glucose Deprivation in HT22 Cells
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HT22 cells were obtained from the American Type Culture Collection (Manassas, VA, USA) and cultured in an incubator with Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 100 U/mL penicillin and streptomycin at 37° C under 5% CO2/95% O2. For oxygen-glucose deprivation (OGD) treatment, HT22 cells were placed first in deoxygenated glucose-free medium and then incubated in a hypoxic vessel for 4 h at 37° C under 95% N2/5% CO2. The cells were then transferred to DMEM supplemented with high glucose and 10% FBS under normoxic conditions (5% CO2) at 37° C for 24 h, as previously described [30 (link)].
10
Culturing Hippocampal Neuronal HT22 Cells
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The hippocampal neuronal HT22 cells of mice were purchased from the American Type Culture Collection (ATCC). HT22 cells were maintained in Dulbecco's modi ed Eagle's medium (Gibco, Grand Island, USA) containing 10% foetal bovine serum (Gibco, Grand Island, USA) and 1% penicillin-streptomycin solution (Life Technologies, Carlsbad, ON, Canada) at 37 °C in a humidi ed incubator with 5% CO 2 . The medium was completely changed every 72 h.
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