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CA-77

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編號(hào):B214889

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產(chǎn)品名稱	CA-77
商品貨號(hào)	B214889
Organism	Rattus norvegicus, rat
Tissue	thyroid C cells
Product Format	frozen
Morphology	neuronal-like
Culture Properties	adherent
Biosafety Level	1 Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Disease	medullary thyroid carcinoma
Applications	neuronal differentiation and peptide secretion studies
Storage Conditions	liquid nitrogen vapor phase
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 ATCC-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. Remove and discard culture medium. 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. 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. Add 5 mL of complete growth medium and aspirate cells by gently pipetting. Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant. 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. 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
Name of Depositor	B Sanborn
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 Birnbaum RS, et al. Identification of procalcitonin in a rat medullary thyroid carcinoma cell line. J. Biol. Chem. 259(5): 2870-4, 1984. PubMed: 6199352 Russo AF, et al. Neuronal properties of a thyroid C-cell line: partial repression by dexamethasone and retinoic acid. Mol. Endocrinol. 6(2): 207-18, 1992. PubMed: 1569964 Birnbaum RS, et al. Biosynthesis of calcitonin by a rat medullary thyroid carcinoma cell line. J. Biol. Chem. 261(2): 699-703, 1986. PubMed: 3941098 Durham PL, et al. Repression of the calcitonin gene-related peptide promoter by 5-HT1 receptor activation. J. Neurosci. 17(24): 9545-53, 1997. PubMed: 9391009 Wood JL, et al. Autoregulation of cell-specific MAP kinase control of the tryptophan hydroxylase promoter. J. Biol. Chem. 276(24): 21262-71, 2001. PubMed: 11283010 Viney TJ, et al. Regulation of the cell-specific calcitonin/calcitonin gene-related peptide enhancer by USF and the Foxa2 forkhead protein. J. Biol. Chem. 279(48): 49948-55, 2004. PubMed: 15385550 Park KY, et al. Control of the calcitonin gene-related peptide enhancer by upstream stimulatory factor in trigeminal ganglion neurons. J. Biol. Chem. 283(9):5 441-51, 2008. PubMed: 18167349

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