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The MB49 cell line is a murine model derived from the C57BL/6 mouse bladder epithelial cells. It was originally developed to study bladder cancer, providing a platform for examining the biological and molecular characteristics of urothelial carcinoma. The cell line was established through the chemical induction of bladder tumors using the carcinogen 7,12-dimethylbenz[a]anthracene (DMBA), as detailed in early research studies. MB49 cells exhibit a tumorigenic phenotype when transplanted into syngeneic mice, forming urothelial carcinomas. These tumors are often poorly differentiated and can display mixed morphologies, including spindle-shaped cells and adenocarcinomatous areas, which resemble aggressive bladder cancer subtypes seen in human pathology.
Further research has led to the development of MB49-I, a more invasive subline of MB49. This subline was generated after 13 consecutive in vivo passages, enhancing its invasive and metastatic potential. MB49-I cells exhibit increased proteolytic activity, particularly in enzymes such as cathepsin B, matrix metalloproteinase 9 (MMP-9), and urokinase-type plasminogen activator (uPA). These enzymes contribute to the breakdown of extracellular matrix components, facilitating the invasion and metastasis of tumor cells. The MB49-I subline, when inoculated orthotopically into the bladder of syngeneic mice, leads to the formation of highly invasive bladder tumors, making it a valuable model for studying tumor progression and testing anti-cancer therapeutics aimed at preventing invasion and metastasis.
This MB49 model, including the MB49-I variant, is instrumental in understanding the molecular mechanisms underlying bladder cancer progression and in developing new therapeutic strategies. The model closely mimics human bladder cancer, particularly in its ability to simulate the invasive and metastatic characteristics of the disease, thereby providing a robust system for preclinical studies.
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Karyotype: Has lost chromosome Y
- 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.
- 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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