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BNL CL.2, a mouse liver cell line originally derived from BALB/c embryonic liver cells, plays a significant role in the study of cellular biology and molecular mechanisms, especially regarding the cell cycle and its regulation. Researchers have extensively used BNL CL.2 to characterize cyclin-dependent kinase (CDK) protein complexes and investigate the alterations in these complexes following both chemical and viral transformation. This line serves as a progenitor for various transformed cell lines such as BNL 1ME A.7R.1, BNL 1NG A.2, and BNL SV A.8, all of which originate from BNL CL.2 and have proven essential for studying CDK alterations post-transformation.
BNL CL.2 is distinguished by its non-tumorigenic nature when tested in immunosuppressed mice, and its inability to grow anchorage-independently, although it does possess the capability to form colonies in semisolid media. This makes it an invaluable model for exploring cellular processes and transformations in a controlled environment. In contrast, its derivative lines such as those transformed by 3-Methylcholanthrene epoxide, MNNG, and SV40 demonstrate the ability to grow in soft agar and form tumors in immunodeficient mice, highlighting the impact of genetic and environmental alterations on cellular behavior. The BNL CL.2 cell line and its derivatives continue to provide a robust foundation for research in cellular transformation, stable cell transfection, and related fields of cellular and molecular biology.
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Tumorigenic: No, the cells were not tumorigenic in immunosuppressed mice, but did form colonies in semisolid medium.
- 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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