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| NCI-H2342 [H2342] cell line Cat No.: NTCC-CRL5941 OrganismHomo sapiens, human Tissuelung Product Formatfrozen Morphologyepithelial Culture Propertiesadherent Biosafety Level1 Diseasestage 3A, adenocarcinoma; non-small cell lung cancer Age55 years Gendermale EthnicityCaucasian DerivationThe line was established in April 1990 from lung adenocarcinoma. Clinical Data55 years Caucasian male The tissue donor was a non-smoker. Complete Growth MediumHITES medium supplemented with 5% fetal bovine serum The base medium for this cell line is DMEM:F12 Medium. To make the complete growth medium,add the following components to the base medium 1.0.005 mg/ml Insulin 2.0.01 mg/ml Transferrin 3.30nM Sodium selenite (final conc.) 4.10 nM Hydrocortisone (final conc.) 5.10 nM beta-estradiol (final conc.) 6.extra 2mM L-glutamine (for final conc. of 4.5 mM) 7.5% fetal bovine serum (final conc.) SubculturingVolumes 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 with 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 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 2.0 to 3.0 mL of complete growth medium and aspirate cells by gently pipetting 5.Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels. 6.Incubate cultures at 37°C. Subcultivation Ratio: 1:2 to 1:6. CryopreservationCulture medium, 95%; DMSO, 5% Culture ConditionsTemperature: 37°C STR ProfileAmelogenin: X,Y CSF1PO: 10 D13S317: 12 D16S539: 13 D5S818: 12 D7S820: 10 TH01: 9,9.3 TPOX: 8,10 vWA: 17 Year of Origin1990 ReferencesNCI-Navy Medical Oncology Branch Cell Line Supplement. J. Cell. Biochem. suppl. 24: 1996. |
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