HT22 Cells
Basic details about the HT-22 neuronal cell line
Description | 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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Organism | Mouse |
Tissue | Brain, hippocampus |
Synonyms | HT-22 |
Aspects
Morphology | Epithelial |
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Growth properties | Adherent |
Documentation
Citation | HT22 (Cytion catalog number 305158) |
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Biosafety level | 1 |
Genetic profile
HT-22 cell culture practices
Culture Medium | DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 1.5 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a) |
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Medium supplements | Supplement the medium with 10% FBS |
Passaging solution | 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. |
Split ratio | 1:3 to 1:6 |
Freeze medium | CM-1 (Cytion catalog number 800100) |
Handling of cryopreserved cultures |
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Quality verification on HT22 cells
Sterility | Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
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Required products
- A Gentle Alternative to Trypsin
Accutase is a cell detachment solution that is revolutionizing the cell culture industry. It is a mix of proteolytic and collagenolytic enzymes that mimics the action of trypsin and collagenase. Unlike trypsin, Accutase does not contain any mammalian or bacterial components and is much gentler on cells, making it an ideal solution for the routine detachment of cells from standard tissue culture plasticware and adhesion coated plasticware. In this blog post, we will explore the benefits and uses of Accutase and how it is changing the game in cell culture.
Advantages of Accutase
Accutase has several advantages over traditional trypsin solutions. Firstly, it can be used whenever gentle and efficient detachment of any adherent cell line is needed, making it a direct replacement for trypsin. Secondly, Accutase works extremely well on embryonic and neuronal stem cells, and it has been shown to maintain the viability of these cells after passaging. Thirdly, Accutase preserves most epitopes for subsequent flow cytometry analysis, making it ideal for cell surface marker analysis.
Additionally, Accutase does not need to be neutralized when passaging adherent cells. The addition of more media after the cells are split dilutes Accutase so it is no longer able to detach cells. This eliminates the need for an inactivation step and saves time for cell culture technicians. Finally, Accutase does not need to be aliquoted, and a bottle is stable in the refrigerator for 2 months.
Applications of Accutase
Accutase is a direct replacement for trypsin solution and can be used for the passaging of cell lines. Additionally, Accutase performs well when detaching cells for the analysis of many cell surface markers using flow cytometry and for cell sorting. Other downstream applications of Accutase treatment include analysis of cell surface markers, virus growth assay, cell proliferation, tumor cell migration assays, routine cell passage, production scale-up (bioreactor), and flow cytometry.
Composition of Accutase
Accutase contains no mammalian or bacterial components and is a natural enzyme mixture with proteolytic and collagenolytic enzyme activity. It is formulated at a much lower concentration than trypsin and collagenase, making it less toxic and gentler, but just as effective.
Efficiency of Accutase
Accutase has been shown to be efficient in detaching primary and stem cells and maintaining high cell viability compared to animal origin enzymes such as trypsin. 100% of cells are recovered after 10 minutes, and there is no harm in leaving cells in Accutase for up to 45 minutes, thanks to autodigestion of Accutase.
In summary
In conclusion, Accutase is a powerful solution that is changing the game in cell culture. With its gentle nature, efficiency, and versatility, Accutase is the ideal alternative to trypsin. If you are looking for a reliable and efficient solution for cell detachment, Accutase is the solution for you.