RBL-2H3大鼠嗜碱性粒细胞 BioVector NTCC质粒载体菌种细胞基因保藏中心

RBL-2H3大鼠嗜碱性粒细胞 BioVector NTCC质粒载体菌种细胞基因保藏中心 RBL-2H3 cell line Cell Type: basophil; chemically induce / Tissue: peripheral blood / Disease: basophilic leukemia Cat No.: NTCC592275 Tissue peripheral blood Cell Type basophil; chemically induce Product Format frozen Morphology fibroblast Culture Properties adherent Biosafety Level 1 Disease basophilic leukemia Strain Wistar Applications This cell line is a suitable transfection host. 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. They have been used extensively to study FcERI and the biochemical pathways for secretion in mast cells. Derivation 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. Receptor Expression FcERI (Fc of IgE) Genes Expressed histamine Cellular Products histamine Comments 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. RBL-2H3 cells have been the model for studies of structure of FcERI. 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 NTCC CRL-1378)that does not degranulate. Histamine release capacity may be seriously reduced after too much subculturing. ref Complete Growth Medium The base medium for this cell line is Eagle's Minimum Essential Medium. To make the complete growth medium, add the following components to the base medium: heat-inactivated fetal bovine serum to a final concentration of 15%. Subculturing Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. 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 Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994. Cryopreservation Freeze medium: Complete growth medium, 95%; DMSO, 5% Storage temperature: liquid nitrogen vapor temperature Culture Conditions Temperature: 37°C Atmosphere: air, 95%; carbon dioxide (CO2), 5% Name of Depositor RP Siraganian References Kulczycki A Jr., et al. The interaction of IgE with rat basophilic leukemia cells. I. Evidence for specific binding of IgE. J. Exp. Med. 139: 600-616, 1974. PubMed: 4812630 Eccleston E, et al. Basophilic leukaemia in the albino rat and a demonstration of the basopoietin. Nat. New Biol. 244: 73-76, 1973. BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心www.biovector.net [Supplier来源] http://www.biovector.net