| Field | Specification |
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The CDNR4 cell line comprises a specialized subset originating from the COMMA-D cell line, known for modeling mouse mammary carcinoma. This clonal subpopulation has undergone extensive characterization, revealing a range of unique properties and functionalities. One of the most striking features of CDNR4 cells is their resemblance to mammary stem cells, which positions them as a significant resource for exploring aspects of stem cell biology, carcinogenesis, and cellular heterogeneity within populations. These cells were developed through the transfection of a transposon bearing kanamycin and neomycin resistance genes (Tn5 gene), which led to the emergence of various intriguing traits and capabilities, including their potential to differentiate into both preneoplastic and neoplastic phenotypes.
Originating from the COMMA-D line, which was initially studied for its cellular heterogeneity using a variety of techniques such as phase contrast microscopy, immunocytochemical staining, DNA content analysis, and evaluations of oncogenic potential, CDNR4 stands out as a distinct clone. Through specific transfection and selection methods, clonal subpopulations like CDNR4 were isolated, each maintaining a degree of the heterogeneity seen in the original COMMA-D parental cells. This retention of heterogeneity underscores the complex nature of these cell populations and enhances the value of CDNR4 cells in research focused on cellular differentiation and cancer progression.
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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
- 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.
- seedingDensity: 2 x 104 cells/cm2 is recommended
- fluidRenewal: 2 to 3 times per week
- postThawRecovery: After thawing, plate the cells at 5 x 104 cells/cm2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours.
- 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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