CT26.CL25 Cells
USD$550.00*
Products are shipped frozen on dry ice in cryotubes. Each cryotube typically contains 3 × 106 cells for adherent lines or 5 × 106 cells for suspension lines (refer to the batch CoA for details).
General information
| Description | 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. |
|---|---|
| Organism | Mouse |
| Tissue | Colon |
| Disease | Adenocarcinoma |
| Synonyms | CT26-clone 25 |
Characteristics
| Breed/Subspecies | BALB/c |
|---|---|
| Age | Unspecified |
| Gender | Female |
| Morphology | Fibroblast |
| Growth properties | Adherent |
Regulatory Data
| Citation | CT26.CL25 (Cytion catalog number 305353) |
|---|---|
| Biosafety level | 1 |
| NCBI_TaxID | 10090 |
| CellosaurusAccession | CVCL_7255 |
| GMO Status | GMO-S1: This murine colon carcinoma cell line (CT26.CL25) contains a retroviral vector encoding lacZ and Tn5-neo, enabling β-galactosidase expression and neomycin resistance. The construct is stably integrated into CT26 cells. This classification applies only within Germany and may differ elsewhere. |
Biomolecular Data
| 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 |
Handling
| Culture Medium | 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 |
| Dissociation Reagent | 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. |
| Freeze medium | As a cryopreservation medium, we 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. |
| Thawing and Culturing Cells |
|
| Incubation Atmosphere | 37°C, 5% CO2, humidified atmosphere. |
| Shipping Conditions | 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. |
| Storage Conditions | 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. |
Quality Control & Molecular Analysis
| Sterility | 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. |
|---|
Certificate of Analysis (CoA)
| Lot Number | Certificate Type | Date | Catalog Number |
|---|---|---|---|
| 305353-200325 | Certificate of Analysis | 23. May. 2025 | 305353 |