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CTLA4 Ig-24 cells, derived from an adult female Chinese hamster (Cricetulus griseus), are a spontaneously immortalized cell line which have been genetically modified by introducing the human CTLA-4 gene, resulting in the expression of a fusion protein.
The fusion protein possesses a dominant attribute of CTLA4Ig, making CTLA4 Ig-24 cells a unique and essential tool in immunology research. CTLA-4, a member of the immunoglobulin superfamily, is primarily expressed in activated T cells and acts as an inhibitory signal transmitter to regulate T cell function.
It shares homology with CD28, a T-cell co-stimulatory protein, and both molecules bind to CD80 (B7-1) and CD86 (B7-2) proteins on antigen-presenting cells. CTLA-4 demonstrates a greater affinity and avidity for CD80 and CD86 than CD28, allowing it to outcompete CD28 for binding to these ligands. By doing so, CTLA-4 transmits an inhibitory signal to T cells, while CD28 transmits a stimulatory signal.
This intricate regulatory mechanism is pivotal in maintaining immune balance and preventing excessive immune responses.
Interestingly, CTLA-4 is also found in regulatory T cells (Tregs) and contributes to their inhibitory function.
When T cells are activated through the T cell receptor (TCR) and CD28, the expression of CTLA-4 increases. Furthermore, CTLA-4 may influence cell motility and signal signalling through the PI3 kinase pathway.
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- 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, 0.2 mM proline, 0.001 mM methotrexate
- 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.
- fluidRenewal: 2 to 3 times per week
- 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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