Large scale isolation of intact rat basophilic leukemia (RBL-2H3) cells.
The rat basophilic leukemia RBL-2H3 mast cell line is widely used for studies of the structure and function of the high affinity IgE receptor (Fc epsilonRI). Here we report on a simple method to isolate large numbers of intact RBL-2H3 cells from tumors produced by injection of the cells into newborn rats. Collagenase treatment of rat tumors yields approximately 3.5 x 10(8) viable cells/animal. Aggregating Fc epsilonRI on these cells induced tyrosine phosphorylation of proteins including the protein tyrosine kinase Syk. This procedure should prove useful for the isolation and characterization of cellular molecules important for mast cell and basophil function.
Hamawy
M M
MM
Laboratory of Immunology, National Institute of Dental Research, National Institutes of Health, Bethesda, MD 20892, USA. hamawy@yoda.nidr.nih.gov
Siraganian
R P
RP
D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D002469:Cell Separation / Q000379:methods; D004792:Enzyme Precursors / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D015471:Leukemia, Basophilic, Acute / Q000473:pathology*; D008407:Mast Cells; D019000:Phosphotyrosine / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D051381:Rats; D017455:Receptors, IgE / Q000502:physiology; D015398:Signal Transduction; D000072377:Syk Kinase
D004792:Enzyme Precursors; D047908:Intracellular Signaling Peptides and Proteins; D017455:Receptors, IgE; D019000:Phosphotyrosine; D011505:Protein-Tyrosine Kinases; D000072377:Syk Kinase; C000606898:Syk protein, rat