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The CHO-CTLA4 cell line is a stable recombinant CHO (Chinese Hamster Ovary) cell line engineered to express the CTLA4 receptor at a medium-low level, approximately 3,000 molecules per cell. This cell line was created using an innovative landing pad technology that facilitates the targeted integration of the CTLA4 gene at a specific, pre-validated genomic locus. CTLA4, also known as CD152, is a critical immune checkpoint protein found primarily on T-cells. It functions by competing with CD28 for binding to B7 molecules (CD80 and CD86) on antigen-presenting cells, leading to the downregulation of T cell activation. This mechanism is vital for maintaining immune self-tolerance and preventing autoimmunity. CTLA4's role in modulating immune responses has made it a significant target in cancer immunotherapy, particularly in immune checkpoint blockade strategies.
The expression of CXCR7 in this cell line was confirmed using flow cytometry.
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Receptors expressed: CTLA4 (CD152)
- cultureMedium: For adherent cultures: 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)For suspension cultures: CHO Growth Medium A (from InSCREENeX; InSCREENeX catalog number INS-ME-1039)
- supplements: For adherent cultures: Supplement the medium with 5% FBS. Add Geneticin (G418-Sulfat) to achieve a final concentration of 0.5 mg/mL.
- dissociationReagent: For adherent cultures: Trypsin-EDTA
- subculturing: For routine adherent cell culture: Aspirate the old culture medium from the adherent cells, and wash them with PBS to remove any remaining medium. After aspirating the PBS, add the appropriate volume of Trypsin/EDTA solution based on the culture vessel size (e.g., 1 ml for a T25 flask, 3 ml for a T75 flask) and incubate at room temperature or 37°C for 5-10 minutes, or until the cells detach. Monitor detachment under a microscope, and gently tap the vessel if necessary to release the cells. Once detached, add complete medium to inactivate the Trypsin/EDTA, gently resuspend the cells, and transfer an aliquot of the cell suspension into a new culture vessel containing fresh medium. Place the vessel in an incubator set to 37°C with 5% CO2, and change the medium every 2-3 days.
- fluidRenewal: 2 to 3 times per week
- postThawRecovery: After thawing, split the cells at a ratio of 1:2 to 1:3 in T25 flasks and allow the cells to recover from the freezing process and to adhere (for adherent cultures) 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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