A549 (ATCC® CRM-CCL-185)

Organism: Homo sapiens, human  /  Cell Type: epithelial cell (KRAS CRM)  /  Tissue: Lung  /  Disease: Carcinoma

Organism Homo sapiens, human
Tissue Lung
Cell Type epithelial cell (KRAS CRM)
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease Carcinoma
Age 58 years
Gender Male
Ethnicity Caucasian
Applications

For use in testing and calibration in ISO 17025 accredited laboratories, to challenge assay performance, validate or compare test methods, and to establish sensitivity, linearity and specificity during assay validation or implementation. ISO Guide 34:2009
 

This cell line may be used as a transfection host.

Intended Use


Certified Reference Material produced under an ISO Guide 34:2009 accredited process. 


    ACLASS Accredited Reference Material Producer AR-1384

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.
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.
Comments

Certificates of Analysis are available electronically at www.atcc.org, or by hardcopy upon request.

Certified Reference Material produced under an ISO Guide 34:2009 accredited process.

This cell line has been tested for the KRAS mutation (p.G12S c.34G>A)

Studies by M. Lieber, et al. revealed that A549 cells could synthesize lecithin with a high percentage of desaturated 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
  1. Remove and discard culture medium.
  2. 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.
  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 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. 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.
  6. 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
STR Profile Amelogenin: X,Y
 CSF1PO: 10, 12
 D13S317: 11
 D16S539: 11,12
 D5S818: 11
 D7S820: 8, 11
 TH01: 8, 9.3 
 TPOX: 8, 11
 vWA: 14
Population Doubling Time about 22 hours
Year of Origin 1972
References

Giard DJ, et al. In vitro cultivation of human tumors: establishment of cell lines derived from a series of solid tumors. J. Natl. Cancer Inst. 51: 1417-1423, 1973. PubMed: 4357758

King BL, et al. Repeat expansion detection analysis of (CAG)n tracts in tumor cell lines, testicular tumors, and testicular cancer families. Cancer Res. 57: 209-214, 1997. PubMed: 9000556

Jeffers M, et al. Degradation of the Met tyrosine kinase receptor by the ubiquitin-proteasome pathway. Mol. Cell. Biol. 17: 799-808, 1997. PubMed: 9001234

Siim BG, et al. Tirapazamine-induced cytotoxicity and DNA damage in transplanted tumors: Relationship to tumor hypoxia. Cancer Res. 57: 2922-2928, 1997. PubMed: 9230202

Zhao Y, et al. Cloning and chromosomal location of a novel member of the myotonic dystrophy family of protein kinases. J. Biol. Chem. 272: 10013-10020, 1997. PubMed: 9092543

Goodrum FD, Ornelles DA. The early region 1B 55-kilodalton oncoprotein of adenovirus relieves growth restrictions imposed on viral replication by the cell cycle. J. Virol. 71: 548-561, 1997. PubMed: 8985383

Putzer BM, et al. Interleukin 12 and B7-1 costimulatory molecule expressed by an adenovirus vector act synergistically to facilitate tumor regression. Proc. Natl. Acad. Sci. USA 94: 10889-10894, 1997. PubMed: 9380730

Pulli T, et al. Cell-surface interactions of echovirus 22. J. Biol. Chem. 272: 21176-21180, 1997. PubMed: 9261123

de Caestecker MP, et al. Characterization of functional domains within Smad4/DPC4. J. Biol. Chem. 272: 13690-13696, 1997. PubMed: 9153220

Li Y, et al. Nuclear factor-kappaB is activated by hyperoxia but does not protect from cell death. J. Biol. Chem. 272: 20646-20649, 1997. PubMed: 9252381

Gao E, et al. The basic helix-loop-helix-zipper transcription factor USF1 regulates expression of the surfactant protein-A gene. J. Biol. Chem. 272: 23398-23406, 1997. PubMed: 9287355

Ray P, et al. Selective up-regulation of cytokine-induced RANTES gene expression in lung epithelial cells by overexpression of IkappaBR. J. Biol. Chem. 272: 20191-20197, 1997. PubMed: 9242696

Shimomura T, et al. Hepatocyte growth factor activator inhibitor, a novel Kunitz-type serine protease inhibitor. J. Biol. Chem. 272: 6370-6376, 1997. PubMed: 9045658

Kumar AS, et al. Phorbol ester down-regulation of lung surfactant protein B gene expression by cytoplasmic trapping of thyroid transcription factor-1 and hepatocyte nuclear factor 3. J. Biol. Chem. 272: 20764-20773, 1997. PubMed: 9252399

Guo FH, et al. Interferon gamma and interleukin 4 stimulate prolonged expression of inducible nitric oxide synthase in human airway epithelium through synthesis of soluble mediators. J. Clin. Invest. 100: 829-838, 1997. PubMed: 9259582

Basic Documentation
FAQ's
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    Date Updated: 1/2/2013

References

Giard DJ, et al. In vitro cultivation of human tumors: establishment of cell lines derived from a series of solid tumors. J. Natl. Cancer Inst. 51: 1417-1423, 1973. PubMed: 4357758

King BL, et al. Repeat expansion detection analysis of (CAG)n tracts in tumor cell lines, testicular tumors, and testicular cancer families. Cancer Res. 57: 209-214, 1997. PubMed: 9000556

Jeffers M, et al. Degradation of the Met tyrosine kinase receptor by the ubiquitin-proteasome pathway. Mol. Cell. Biol. 17: 799-808, 1997. PubMed: 9001234

Siim BG, et al. Tirapazamine-induced cytotoxicity and DNA damage in transplanted tumors: Relationship to tumor hypoxia. Cancer Res. 57: 2922-2928, 1997. PubMed: 9230202

Zhao Y, et al. Cloning and chromosomal location of a novel member of the myotonic dystrophy family of protein kinases. J. Biol. Chem. 272: 10013-10020, 1997. PubMed: 9092543

Goodrum FD, Ornelles DA. The early region 1B 55-kilodalton oncoprotein of adenovirus relieves growth restrictions imposed on viral replication by the cell cycle. J. Virol. 71: 548-561, 1997. PubMed: 8985383

Putzer BM, et al. Interleukin 12 and B7-1 costimulatory molecule expressed by an adenovirus vector act synergistically to facilitate tumor regression. Proc. Natl. Acad. Sci. USA 94: 10889-10894, 1997. PubMed: 9380730

Pulli T, et al. Cell-surface interactions of echovirus 22. J. Biol. Chem. 272: 21176-21180, 1997. PubMed: 9261123

de Caestecker MP, et al. Characterization of functional domains within Smad4/DPC4. J. Biol. Chem. 272: 13690-13696, 1997. PubMed: 9153220

Li Y, et al. Nuclear factor-kappaB is activated by hyperoxia but does not protect from cell death. J. Biol. Chem. 272: 20646-20649, 1997. PubMed: 9252381

Gao E, et al. The basic helix-loop-helix-zipper transcription factor USF1 regulates expression of the surfactant protein-A gene. J. Biol. Chem. 272: 23398-23406, 1997. PubMed: 9287355

Ray P, et al. Selective up-regulation of cytokine-induced RANTES gene expression in lung epithelial cells by overexpression of IkappaBR. J. Biol. Chem. 272: 20191-20197, 1997. PubMed: 9242696

Shimomura T, et al. Hepatocyte growth factor activator inhibitor, a novel Kunitz-type serine protease inhibitor. J. Biol. Chem. 272: 6370-6376, 1997. PubMed: 9045658

Kumar AS, et al. Phorbol ester down-regulation of lung surfactant protein B gene expression by cytoplasmic trapping of thyroid transcription factor-1 and hepatocyte nuclear factor 3. J. Biol. Chem. 272: 20764-20773, 1997. PubMed: 9252399

Guo FH, et al. Interferon gamma and interleukin 4 stimulate prolonged expression of inducible nitric oxide synthase in human airway epithelium through synthesis of soluble mediators. J. Clin. Invest. 100: 829-838, 1997. PubMed: 9259582