| Field | Specification |
|---|---|
| Species |
The C6 cell line maintains glial cell type with fibroblast morphology and originates from a glioma of a Wisthar-Furth rat. The glioma was induced by exposure to N-nitrosomethylurea, following numerous cycles of alternating culture and animal passages.
The C6 glioma cell line is frequently utilized in neuro-oncology research to create animal models that closely mimic the characteristics of human glioma, aiding in the development of new therapeutic agents and strategies. It is particularly effective in 3D cell culture and high-throughput screening.
C6 cells are genetically diverse, possessing a wild-type p53 gene, increased Rb gene expression, and a mutant p16/Cdkn2a/Ink4a locus but lacking p16 and p19ARF mRNA expression. They also overexpress several genes in human gliomas, such as PDGFβ, IGF-1, EGFR, and Erb3/Her3 precursor proteins.
However, the expression of IGF-2, FGF-9, and FGF-10 is reduced, while MMP-7 gene expression remains unchanged. Like human gliomas, C6 cells show increased activity of the Ras pathway genes, which is regulated by the elevated expression of the Ras guanine triphosphate activator protein.
The C6 cell line has been utilized in various studies. For instance, it was used to examine the ability of 2-(2,4-dihydroxy phenyl)thieno-1,3-thiazin-4-one (BChTT) to halt cancer cell proliferation and to investigate the mechanisms involved in this process.
In another research, the cytotoxic and antioxidant properties of the supercritical CO2 extract (SCE) of an old man's beard (Usnea barbata) were studied using C6 cells. Interestingly, these cells have been reported to show increased levels of glyceryl phosphate dehydrogenase activity in response to glucocorticoids.
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- Receptors expressed: Glucocorticoid
- Viruses: Positive for LCMV
- Virus susceptibility: vesicular stomatitis (Indiana), vaccinia, herpes simplex
- Virus resistance: poliovirus 3
- Reverse transcriptase: negative
- Products: S-100 protein, production of glyceryl phosphate dehydrogenase in response to glucocorticoids, somatotrophin.
- cultureMedium: RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- doublingTime: 24 hours
- 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.
- seedingDensity: 1 x 104 cells/cm2 will yield in a confluent layer in about 4 days
- fluidRenewal: 2 to 3 times per week
- postThawRecovery: After thawing, plate the cells at 5 x 104 cells/cm2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours.
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, 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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