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NCI-H1869; Lung Carcinoma; Human (Homo sapiens) Item ID: ATCCRL5900

Permits and Restrictions

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Organism Homo sapiens, human
Tissue lung
Cell Type squamous cell
Product Format frozen
Culture Properties adherent, 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 stage 4, non-small cell lung cancer
Age 58 years
Gender male
Ethnicity Caucasian
Storage Conditions liquid nitrogen vapor phase
Derivation The line was established in March 1988 from pleural effusion, a metastatic site.
Clinical Data The patient received prior chemotherapy and radiation therapy. The patient was a smoker. 50 pack years.
Complete Growth Medium ACL-4 medium supplemented with 10% FBSThe 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:
  1. 0.02 mg/ml insulin
  2. 0.01 mg/ml transferrin
  3. 25 nM sodium selenite
  4. 50 nM Hydrocortisone
  5. 1 ng/ml Epidermal Growth Factor (do not filter)
  6. 0.01 mM ethanolamine
  7. 0.01 mM phosphorylethanolamine
  8. 100 pM triiodothyronine
  9. 0.5% (w/v) bovine serum albumin
  10. 10 mM HEPES
  11. 0.5 mM sodium pyruvate
  12. 2mM L-glutamine(for final conc. of 4.5 mM)
  13. 10% FBS
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.
  1. Remove and discard culture medium.
  2. 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.
  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.
  6. Incubate cultures at 37°C.

Subcultivation Ratio: 1:4 to 1:6Medium Renewal: Every 2 to 3 days

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation RPMI 1640 medium 85%; fetal bovine serum, 10%; DMSO, 5%. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.
Culture Conditions Temperature: 37°C
STR Profile Amelogenin: X CSF1PO: 10 D13S317: 11 D16S539: 12 D5S818: 11 D7S820: 8,9 THO1: 6 TPOX: 8,9 vWA: 16
Name of Depositor AF Gazdar, JD Minna
Year of Origin 1988
References

NCI-Navy Medical Oncology Branch Cell Line Supplement. J. Cell. Biochem. suppl. 24: 1996.

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

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