CT26.CL25

The CT26.CL25 cell line is a murine colon carcinoma model derived from the parental CT26 cell line, which is a chemically induced, undifferentiated colon carcinoma originating from BALB/c mice. CT26.CL25 has been genetically modified to express the β-galactosidase (β-gal) protein, making it an excellent model for studying tumor immunology and immunotherapy, particularly in the context of tumor-associated antigens (TAAs). This modification allows for specific immunological studies targeting β-gal as a neoantigen, facilitating research into the mechanisms of tumor immune evasion and the development of cancer vaccines or adoptive cell therapies.

CT26.CL25 has been employed in preclinical models to investigate immune responses and the efficacy of immunotherapies, such as the use of dendritic cells (DCs) loaded with tumor-associated antigens. Studies have shown that immunization strategies using DCs pulsed with peptides derived from retroviral antigens, like gp70, can elicit robust anti-tumor immune responses. In experimental models, the activation of CD8+ cytotoxic T lymphocytes (CTLs) specific for gp70 was observed, demonstrating the cell line's utility in testing immunotherapeutic approaches. However, the immunization with such peptide-loaded DCs has shown limitations, particularly in treating established metastases, highlighting the challenges in translating prophylactic immune responses into therapeutic efficacy.

Additionally, CT26.CL25 is often used in research to test the efficacy of combined immunotherapy approaches, such as the use of immune checkpoint inhibitors or cancer vaccines. For instance, studies have evaluated the impact of metronomic chemotherapy combined with immune checkpoint inhibitors, where the induction of immunogenic cell death (ICD) in CT26.CL25 has been crucial for enhancing the anti-tumor immune response. These investigations have demonstrated that targeting immune checkpoints can synergize with chemotherapy to increase tumor rejection rates and establish long-term immunological memory.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Mycoplasma contamination is 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.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.