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

Organism	Homo sapiens, human
Tissue	lung
Cell Type	epithelial
Product Format	frozen
Morphology	epithelial-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	Carcinoma
Age	58 years
Gender	male
Ethnicity	Caucasian
Applications	This cell line is a suitable transfection host.
Storage Conditions	liquid nitrogen vapor phase

Karyotype	This is a hypotriploid human cell line with the modal chromosome number of 66, occurring in 24% of cells. Cells with 64 (22%), 65, and 67 chromosome counts also occurred at relatively high frequencies; the rate with higher ploidies was low at 0.4%. There were 6 markers present in single copies in all cells. They include der(6)t(1;6) (q11;q27); ?del(6) (p23); del(11) (q21), del(2) (q11), M4 and M5. Most cells had two X and two Y chromosomes. However, one or both Y chromosomes were lost in 40% of 50 cells analyzed. Chromosomes N2 and N6 had single copies per cell; and N12 and N17 usually had 4 copies. Note: Cytogenetic information is based on initial seed stock at ATCC. Cytogenetic instability has been reported in the literature for some cell lines.
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Derivation	This line was initiated in 1972 by D.J. Giard, et al. through explant culture of lung carcinomatous tissue from a 58-year-old Caucasian male.
Antigen Expression	The cells are positive for keratin by immunoperoxidase staining.
Comments	Studies by M. Lieber, et al. revealed that A549 cells could synthesize lecithin with a high percentage of disaturated fatty acids utilizing the cytidine diphosphocholine pathway.

Complete Growth Medium	The base medium for this cell line is ATCC-formulated F-12K Medium, Catalog No. 30-2004. 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 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product. Remove and discard culture medium. 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. 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. Cultures can be established between 2 x 103 and 1 x 104 viable cells/cm2. Do not exceed 7 x 104 cels/cm2. Incubate cultures at 37°C. Interval: Maintain cultures at a cell concentration between 6 X 103 and 6 X 104 cell/cm2. Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:8 is recommended Medium Renewal: 2 to 3 times per week
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