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The GCT cell line, originating from a giant cell tumor (GCT) isolated from the lung of an adult male patient with fibrous histiocytoma, is renowned for its robust biological activity in the field of medical research. This line produces Colony Stimulating Activity (CSA) for human granulocyte precursors and Erythropoietin-like Erythroid Activity (EEA) for erythroid precursors, making it invaluable for studying the regulation and development of hematopoietic cells. The granulocyte and erythroid precursors targeted by the GCT cell line's products are key to understanding processes like neutrophil function in the immune response and red blood cell formation, respectively.
Additionally, the medium conditioned by this cell line is a significant source of prostaglandin E and plasminogen activator. These substances have crucial roles in inflammatory responses and the fibrinolytic pathway, respectively. Prostaglandin E is essential for inflammatory modulation and maintaining physiological balance, while plasminogen activator contributes to the dissolution of blood clots. The presence of these factors in the GCT cell line's conditioned medium underscores its potential for developing therapeutic strategies addressing cardiovascular diseases and conditions related to excessive clot formation and inflammation.
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- cultureMedium: McCoys 5a, w: 3.0 g/L Glucose, w: stable Glutamine, w: 2.0 mM Sodium pyruvate, w: 2.2 g/L NaHCO3 (Cytion article number 820200a)
- 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.
- seedingDensity: 1 to 2 x 104 cells/cm2
- 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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