B16 Cells
Product number:
305154
Key facts about B16 cells
Description | B16 melanoma is a well-established and widely used cell line in cancer research, which was first established in 1954. B16 cells are derived from the melanin-producing epithelia of C57BL/6 mice and are known for their ability to form metastases in the lung, liver, and spleen. By utilizing B16 cells as a research model, researchers were able to gain a deeper understanding of the mechanisms by which cancer cells spread and establish themselves in various body organs. B16 cells express very low amounts of the H-2Kb and H-2Db glycoproteins of mouse class I major histocompatibility complex. Treatment with gamma interferon increases the expression of H-2 and the metastatic potential of B16 cells. This discovery further advanced the understanding of the mechanisms of metastasis. Current research with B16 cells focuses on the immune response of cells to vaccinations, the role of microRNA in modulating metastatic characteristics, and the use of B16 as a preclinical model for researching immunotherapy. There are distinct sub-lines of B16 cells, each with its own particular properties. B164A5 is the most aggressive melanoma cell line for the skin of C57BL/6J mice, followed by B16F10, B16-GMCSF, and B16FLT3. The base medium for B16 cells is DMEM. |
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Organism | Mouse |
Tissue | Skin |
Disease | Mouse melanoma |
Synonyms | B-16, B16 melanoma, B16 subline B78, B78 |
Aspects
Gender | Male |
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Morphology | Mixture of spindle-shaped and epithelial-like cells |
Growth properties | Adherent |
Identifiers / Biosafety / Citation
Citation | B16 (Cytion catalog number 305154) |
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Biosafety level | 1 |
Genetic profile of the mouse melanoma cancer cell line B16
Tumorigenic | Yes |
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Handling B16 cells
Culture Medium | EMEM, w: 2 mM L-Glutamine, w: 1.5 g/L NaHCO3, w: EBSS, w: 1 mM Sodium pyruvate, w: NEAA (Cytion article number 820100c) |
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Medium supplements | Supplement the medium with 10% FBS |
Passaging solution | 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. |
Split ratio | 1:4 to 1:8 |
Fluid renewal | 2 to 3 times per week |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures | B16 cells are shipped in a deep-frozen state on dry ice. Upon receipt, confirm that the vial remains frozen. For storage, place the cryovial immediately at temperatures below -150 degrees. If you plan to culture the cells immediately, swiftly thaw the vial by shaking it in a 37 degrees water bath with clean water and an antimicrobial agent for 40-60 seconds. Remove the vial once a small ice clump persists, ensuring it remains cold. Proceed with all subsequent steps under aseptic conditions. In a sterile flow hood, disinfect the cryovial with 70% ethanol. Then, gently open the vial and transfer the cell suspension into a 15 ml centrifuge tube pre-filled with 8 ml of room temperature culture medium. Gently mix the cells. For cell separation, centrifuge at 300 x g for 3 minutes and dispose of the supernatant. Skipping centrifugation is optional, although any residual freezing medium should be removed after 24 hours. Resuspend the pellet gently in 10 ml of fresh culture medium and divide between two T25 culture flasks. Follow the subculture protocol for subsequent steps. |
Quality Verification
Sterility | Mycoplasma contamination is rigorously 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. |
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