NCI-H1435 [H1435] (ATCC® CRL-5870)

Organism: Homo sapiens, human  /  Tissue: lung  /  Disease: adenocarcinoma; non-small cell lung cancer

Permits and Restrictions

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Organism Homo sapiens, human
Tissue lung
Product Format frozen
Culture Properties adherent, single cells and loosely attached clusters
Biosafety Level 1
Disease adenocarcinoma; non-small cell lung cancer
Age 35 years
Gender female
Storage Conditions liquid nitrogen vapor phase
Derivation
The line was established in June 1986.
Clinical Data
35 years old
female
The tissue donor was a non-smoker.
Comments The tissue donor was a non-smoker.
Complete Growth Medium ACL-4 medium (serum-free)
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:
  • 0.02 mg/ml insulin
  • 0.01 mg/ml transferrin
  • 25 nM sodium selenite (final conc.)
  • 50 nM Hydrocortisone (final conc.)
  • 1 ng/ml Epidermal Growth Factor (do not filter)
  • 0.01 mM ethanolamine (final conc.)
  • 0.01 mM phosphorylethanolamine (final conc.)
  • 100 pM triiodothyronine (final conc.)
  • 0.5% (w/v) bovine serum albumin (final conc.)
  • 10 mM HEPES
  • 0.5 mM sodium pyruvate (final conc.)
  • extra 2mM L-glutamine (for final conc. of 4.5mM)

Subculturing

Volumes 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.
  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. Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant.
  5. To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes.
  6. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
  7. Place culture vessels in incubators at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended
Medium Renewal: Every 2 to 3 days

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Animal Cells: A Manual Of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005.

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: 13
D13S317: 8,11
D16S539: 11
D5S818: 11
D7S820: 12,14
THO1: 6,7
TPOX: 8
vWA: 15,17
Name of Depositor AF Gazdar, JD Minna
Year of Origin June, 1986
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.

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Restrictions

The line is available with the following restrictions: 

  1. This cell line was deposited at the ATCC by Dr. A. Gazdar and Dr. J. Minna and is provided for research purposes only. Neither the cell line nor products derived from it may be sold or used for commercial purposes. Nor can the cells be distributed to third parties for purposes of sale, or producing for sale, cells or their products. The cells are provided as service to the research community. They are provided without warranty of merchantability or fitness for a particular purpose or any other warranty, expressed or implied. 
  2. Any proposed commercial use of the these cells, or their products must first be negotiated with the University of Texas Southwestern Medical Center at Dallas, 5323 Harry Hines Blvd., Dallas, Texas 75235. Telephone (214) 699-8056, FAX (214) 688-7233.

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.