RBL-2H3 Cells

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.The role of calcium in RBL-2H3 cell degranulation is significant. The calcium ionophore A23187, which increases intracellular calcium levels, induces degranulation in RBL-2H3 cells, similar to mast cells and basophils. Some studies have described RBL-2H3 cells as a serotonin-releasing cell line.

Adherent

Wistar

1

Supplement the medium with 10% FBS

2 to 3 times per week

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

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.

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.

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

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.

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

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.

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

When planning to store a cryovial in liquid nitrogen for future thawing, it is mandatory to adhere to stringent safety measures. Appropriate protective gloves and clothing are essential, and the use of a face mask or safety goggles is required during the transfer of frozen samples to or from the liquid nitrogen tank. This is to mitigate the risk of injury from potential cryovial explosions upon removal, which can result in the projection of sharp fragments.

We stand by the promise of delivering products with high cell viability and robust culture performance. To achieve the best results, please make sure you follow the storage and culture instructions detailed in the product information sheet closely. Your adherence to these guidelines is key to success.