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The MNNG/HOS Cl #5 cell line [R-1059-D] is derived from the human osteosarcoma cell line HOS through in vitro transformation with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) at a concentration of 0.01 mcg/ml. This compound is a potent carcinogen, and the transformation resulted in significant tumorigenic properties, evidenced by the formation of tumors in nude mice within 21 days at a 100% frequency when inoculated subcutaneously with 107 cells. These tumors were observed to be poorly differentiated sarcomas or osteosarcomas. The cell line was originally established from a 13-year-old White female patient with osteosarcoma and exhibits adherent growth properties. Functionally, MNNG/HOS Cl #5 cells demonstrate high saturation density and high plating efficiency in soft agar, reflecting their enhanced anchorage-independent growth, a hallmark of malignant transformation. Additionally, these cells exhibit notable fibrinolytic activity, which has been associated with increased tumorigenic potential. When compared to untreated HOS cells, MNNG-treated cells exhibit more robust cell aggregation properties and a higher propensity to form colonies in soft agar, which correlates with their tumor-forming abilities. In experiments, MNNG-transformed cells produced tumors in both nude mice and hamsters, with cells resembling the parent HOS line, while untreated cells were non-tumorigenic under similar conditions. This cell line is also useful in studying cancer progression and tumor biology, particularly osteosarcoma, as it provides a model of chemically induced transformation. The ability of these cells to grow in an immunocompromised environment (e.g., nude mice) makes them a valuable tool for preclinical cancer research, allowing for the investigation of tumorigenic mechanisms and the potential testing of therapeutic interventions.
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- Isoenzymes: G6PD, B
- Tumorigenic: Yes, in nude mice
- cultureMedium: RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a)
- 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: 1 x 104 cells/cm2
- 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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