CTLA4 Ig-24 Cells
Product number:
305014
General information
Description | 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. |
---|---|
Organism | Hamster |
Tissue | Ovary |
Synonyms | CTLA4Ig-24 |
Characteristics
Gender | Female |
---|---|
Morphology | Epithelial |
Growth properties | Adherent |
Identifiers / Biosafety / Citation
Citation | CTLA4 Ig-24 (Cytion catalog number 305014) |
---|---|
Biosafety level | 1 |
Expression / Mutation
Handling
Culture Medium | DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 1.5 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a) |
---|---|
Medium supplements | Supplement the medium with 10% FBS, 0.2 mM proline, 0.001 mM methotrexate |
Passaging solution | 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. |
Split ratio | 1: 3 to 1: 4 |
Fluid renewal | 2 to 3 times per week |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures | CTLA4 Ig-24 cells are shipped in a deep-frozen state on dry ice. Upon receipt, confirm that the vial remains frozen. For storage, place the cryovial immediately at temperatures below -150 degrees. If you plan to culture the cells immediately, swiftly thaw the vial by shaking it in a 37 degrees water bath with clean water and an antimicrobial agent for 40-60 seconds. Remove the vial once a small ice clump persists, ensuring it remains cold. Proceed with all subsequent steps under aseptic conditions. In a sterile flow hood, disinfect the cryovial with 70% ethanol. Then, gently open the vial and transfer the cell suspension into a 15 ml centrifuge tube pre-filled with 8 ml of room temperature culture medium. Gently mix the cells. For cell separation, centrifuge at 300 x g for 3 minutes and dispose of the supernatant. Skipping centrifugation is optional, although any residual freezing medium should be removed after 24 hours. Resuspend the pellet gently in 10 ml of fresh culture medium and divide between two T25 culture flasks. Follow the subculture protocol for subsequent steps. |
Quality control / Genetic profile / HLA
Sterility | Mycoplasma contamination is rigorously excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
---|