FcERI (Fc of IgE)

RBL-2H3 is a basophilic leukemia cell line isolated and cloned in 1978 in the Laboratory of Immunology at the National Institute of Dental Research from Wistar rat basophilic cells that were maintained as tumors. [22638]
These cells have high affinity IgE receptors.
They can be activated to secrete histamine and other mediators by aggregation of these receptors or with calcium ionophores.
They have been used extensively to study FcERI and the biochemical pathways for secretion in mast cells.
RBL-2H3 cells have been the model for studies of structure of FcERI.
They have been used extensively for studies of different aspects of secretion in cells including the role of changes in intracellular calcium, the activation of phospholipases, protein kinases and small G proteins.
Although nearly all lots of fetal bovine serum support the growth of these cells, the cells grown in some lots degranulate better after FcERI aggregation.
Another rat basophil line is available (RBL-1, see ATCC CRL-1378) that does not degranulate.

Protocol:

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
3. Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels.
6. Incubate cultures at 37°C.
   
   
   Subcultivation ratio: A subcultivation ratio of 1:4 to 1:8 is recommended
   
   Medium renewal: Every 2 to 3 days
   

Recommended medium (without the additional supplements or serum described under ATCC Medium): ATCC 30-2003