大鼠甲状腺髓样癌细胞CA77 cell line细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心

大鼠甲状腺髓样癌细胞CA77 cell line细胞株

CA-77 Cell Line Cat No.: NTCC-A3234 Organism Rattus norvegicus, rat Tissue thyroid C cells Product Format frozen Morphology neuronal-like Culture Properties adherent Biosafety Level 1 Disease medullary thyroid carcinoma Applications neuronal differentiation and peptide secretion studies Derivation CA77 was subcloned from a serially passaged collagenase and trypsin digestion of a transplantable rat medullary thyroid carcinoma of the CA tumor series, serum free media was used to restrict fibroblast outgrowth, after 23 subcultures serum was added to media Genes Expressed calcitonin gene-related protein (CGRP) Comments CA77 cells have a neuronal-like phenotype, neurite extension is enhanced by growth on laminin substrate. Cells have a neural crest derived lineage, which makes them a useful model for differentiation studies; expresses calcitonin gene-related peptide (CGRP) and other neuropeptides, which along with the serotonergic neuronal-like phenotype, makes them a useful model for gene expression and other molecular studies. Complete Growth Medium The base medium for this cell line is NTCC-formulated DMEM:F12 Medium Catalog No. 30-2006. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%. Subculturing Volumes used in this protocol are for 75 cm2 flasks; 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, twice, with 5 mL Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor. 3.Add 5 mL of 0.25% 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 5 mL of complete growth medium and aspirate cells by gently pipetting. 5.Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant. 6.Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new vessels. An inoculum of 7.0 X 14 to 1.0 X 105 viable cells/cm2 is recommended. 7.Incubate cultures at 37°C. Subcultivation ratio: A subcultivation ratio of 1:3 to 1:5 is recommended. Medium renewal: Every 2 to 3 days Cryopreservation Complete growth medium, 95% supplemented with 5% DMSO Culture Conditions Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C Year of Origin 1980 References Muszynski M, et al. Glucocorticoids stimulate the production of preprocalcitonin-derived secretory peptides by a rat medullary thyroid carcinoma cell line. J. Biol. Chem. 258(19): 11678-83, 1983. PubMed: 6311819

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