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CTX TNA2 is a rat astrocyte cell line that was established from primary cultures of cortical astrocytes. It is often used to study central nervous system (CNS) functions, particularly in relation to glial biology, neurotoxicity, and neuroprotection. Astrocytes play a critical role in maintaining CNS homeostasis, providing structural and metabolic support to neurons, and mediating responses to injury and oxidative stress.
In various studies, CTX TNA2 cells have been employed to model neurotoxicity, especially involving excitotoxicity induced by agents such as glutamate. For instance, exposure to glutamate in CTX TNA2 cells triggers apoptosis and autophagy through mechanisms involving reactive oxygen species (ROS) and the glycogen synthase kinase-3β (GSK-3β) pathway. These pathways are central to the cells' response to oxidative stress and mitochondrial dysfunction, particularly after traumatic brain injury or other neurodegenerative conditions. Additionally, neuroprotective agents like resveratrol and cannabidiol (CBD) have been shown to reduce ROS generation and inhibit glutamate-induced autophagy and apoptosis in these astrocytes.
The CTX TNA2 cell line has proven to be a valuable in vitro model for studying not only basic astrocyte function but also the therapeutic potential of antioxidant and neuroprotective compounds under conditions of CNS injury and disease.
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Viruses: Transformant: Simian virus 40 (SV40)
- 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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