RBL-2H3 Cells
General information
Description | The RBL-2H3 cell line has become a valuable tool for studying mast cell physiology. RBL-2H3 cells express rat mast cell protease II (RMCP-II) and the c-kit receptor tyrosine kinase, making them a potential model for mast cells. However, conflicting and sometimes misleading data about RBL-2H3 cells have been reported. RBL-2H3 cells have been widely used to investigate various aspects of mast cell function, including degranulation, mast cell stabilizers, and the interaction of FceRI receptors with the cytoskeleton. They express high-affinity IgE receptors and can be activated to secrete histamine and other mediators. Cultivating RBL-2H3 cells is relatively easy, and longer culturing times result in higher cell density. Degranulation is a key feature of RBL-2H3 cells, similar to mast cells and basophils. When allergens crosslink their IgE-bound FceRI receptors, RBL-2H3 cells release preformed and newly synthesized mediators, contributing to immune allergic responses. The degranulation of RBL-2H3 cells has provided insights into basophil degranulation as well. These cells can also undergo degranulation in response to non-immunological stimuli, and there are differences between MMC, RBL-2H3, and CTMC. |
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Organism | Rat |
Tissue | Peripheral blood |
Disease | Rat leukemia |
Synonyms | RBL2H3, RBL 2H3, RBL.2H3 |
Characteristics
Morphology | Fibroblast |
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Growth properties | Adherent |
Identifiers / Biosafety / Citation
Citation | RBL-2H3 (Cytion catalog number 305194) |
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Biosafety level | 1 |
Expression / Mutation
Handling
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 | 1:2 to 1:4 |
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 |
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Quality control / Genetic profile / HLA
Sterility | 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. |
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