MB49

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.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

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.

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.

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.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.