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The NCH640 cell line is a glioblastoma stem-like cell model used in research to explore the mechanisms of tumor resistance, cellular survival under stress, and therapeutic responses. Glioblastoma, one of the most aggressive forms of brain tumors, is difficult to treat due to its resistance to therapy and adaptation to a hostile microenvironment. NCH640 is cultured in specialized media such as Neurobasal A with supplements like B27, and its growth is supported by essential growth factors like EGF and FGF-2. It is often used alongside other glioma stem cell models, such as NCH690 and NCH644, to investigate these biological phenomena.
The research on NCH640 focuses heavily on its resistance mechanisms, particularly under hypoxic conditions. Glioma cells like NCH640 show significant reliance on metabolic adaptations, including altered reactive oxygen species (ROS) regulation. Studies have demonstrated that targeting pathways such as the integrated stress response (ISR) in NCH640 and related cell lines may improve their sensitivity to therapies like temozolomide, which is commonly used in glioblastoma treatment. These findings are important for devising new strategies to overcome the inherent resistance of glioma stem-like cells to standard therapeutic interventions.
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Tumorigenic: Yes
- cultureMedium: DMEM:Ham's F12 (1:1), w: 3.1 g/L Glucose, w: 2.5 mM L-Glutamine, w: 15 mM HEPES, w: 0.5 mM Sodium pyruvate, w: 1.2 g/L NaHCO3 (Cytion article number 820400a)
- supplements: Supplement the medium with 10% FBS, 5 mg/L Heparin, 20 ng/mL bFGF, 20 microgram/L EGF, 5 mg/L Insulin, 100 mg/L Transferrin, 5,2 microgram/L Na-selenit, 6,3 microgram/L Progesteron, 161,1 microgram/L Putrescin, 50 mg/L Hydrocortinson
- subculturing: For subculturing spheroid cultures, begin by mechanically dissociating the spheroids through pipetting up and down 5 to 10 times using an Eppendorf pipette with 1000 μl filter tips. After this, centrifuge the mixture at 300g for 5 minutes at room temperature to pellet the cells. Discard the supernatant and resuspend the cell pellet in fresh culture medium. Finally, transfer the resuspended cells into new culture vessels to promote further spheroid formation. This approach ensures efficient spheroid breakdown and readies them for continued growth in a new environment.
- seedingDensity: 1 x 105 cells/mL
- fluidRenewal: 2 to 3 times per week
- postThawRecovery: After thawing allow the cells to recover from the freezing process for at least 24 to 48 hours.
- 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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