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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.
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- Antigen expression: H-2d
- Tumorigenic: Yes, in BALB/c mice
- Products: Genes expressed: beta galactosidase (beta-gal), H-2D
- Mutational profile: Gene deletion: Cdkn2a, homozygous; Mutation: Kras, p.Gly12Asp (c.35G>A), homozygous
- cultureMedium: RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a)
- supplements: Supplement the medium with 10% FBS, 1% NEAA, 0.4 mg/mL G418, add 2.5 g/L glucose and 10 mM HEPES
- 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.
- 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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