Human Cell Lines-Cell Lines-InCellGene LLC.-CellLines,Biomedicine reasearch Product,Cell Biology,Cell Culture,293T Cell,Media

Permits and Restrictions	View Restrictions
Organism	Homo sapiens, human
Tissue	lung; derived from metastatic site: pleural effusion
Product Format	frozen
Morphology	epithelial
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	adenocarcinoma; non-small cell lung cancer
Age	65 years adult
Gender	male
Ethnicity	Caucasian
Storage Conditions	liquid nitrogen vapor phase

Karyotype	Modal number = 79; range = 71 to 83 This is a hypertriploid human cell line with the modal chromosome number of 79, occurring in 20% of cells. The rate of cells with a higher ploidy was 5.5%. Karyotypes were very complex. There were over 22 marker chromosomes commonly present in most cells, many with complex structural rearrangements. Among these markers were: double copies for t(10qter--10q11.2::?::13C--13qter) and der(9)t(3;9) (p12;p22?), and one copy each for del (1) (p22) and i (iq). There were two normal X chromosomes per cell. Normal chromosomes Y, N1, N5, N14, and N15 were not found. Chromosomes N20 and N22 generally had four or more copies per cell.
Clinical Data	65 years adult Caucasian male
HeLa Markers	N
Tumorigenic	Yes
Effects	Yes, in nude mice (Tumors developed within one month in 4/5 nude mice inoculated subcutaneously with 10(7) cells.)
Comments	An EBV-transformed lymphoblastoid cell line from the same patient is available as ATCC CCL-256.1.

Complete Growth Medium	HITES medium supplemented with 5% fetal bovine serum 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 0.005 mg/ml Insulin 0.01 mg/ml Transferrin 30nM Sodium selenite (final conc.) 10 nM Hydrocortisone (final conc.) 10 nM beta-estradiol (final conc.) extra 2mM L-glutamine (for final conc. of 4.5 mM) 5% fetal bovine serum (final conc.)
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. Remove and discard culture medium. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor. 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. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 6 x 103 to 8 x 103 viable cells/cm2 is recommended. Or use Subcultivation Ratio: 1:2 to 1:4 Incubate cultures at 37°C. Subculture when reaching a cell concentration between 8 x 104 and 1 x 105 cells/cm2. Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended Medium Renewal: Every 2 to 3 days
Cryopreservation	Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase
Culture Conditions	Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C