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C127 cells, originating from murine mammary epithelial tissues, are an indispensable mammalian cell line that lays a solid groundwork for a multitude of biological studies. These cells have undergone a rigorous engineering process, involving infection with specifically designed viruses that integrate T7 RNA polymerase driven by a viral promoter into their genome. The flexibility of C127 cells is further enhanced by the introduction of an additional recombinant virus that carries cystic fibrosis transmembrane conductance regulator (CFTR) cDNA under the control of a T7 promoter, or alternatively, a transfected plasmid bearing the same promoter. This genetic setup enables precise control over protein expression, tailored to produce specific proteins, thereby making C127 cells an exceptional tool for protein expression studies.
The epithelial nature of C127 cells, reflective of their derivation from mammary gland tissues, supports their growth in an adherent manner. They exhibit rapid proliferation and can be employed to scrutinize cellular processes, growth, and differentiation across diverse experimental conditions. The unique genetic modifications present in these cells make them an ideal model for stable cell transfection experiments, allowing researchers to insert foreign genetic material and explore gene functions, protein interactions, and the consequences of genetic modifications. Additionally, their use in 3D cell culture has been increasingly recognized, providing insights into cell-cell interactions, tissue morphogenesis, and disease modeling with greater physiological relevance, thereby extending their utility beyond traditional 2D cultures.
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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.
- fluidRenewal: 2 to 3 times per week
- 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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