NCI-H2029 [H2029] (ATCC® CRL-5913)

Organism: Homo sapiens, human  /  Tissue: lung  /  Disease: stage E, carcinoma; small cell lung cancer

Permits and Restrictions

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Organism Homo sapiens, human
Tissue lung
Product Format frozen
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 stage E, carcinoma; small cell lung cancer
Age 69 years
Gender female
Ethnicity Caucasian
Storage Conditions liquid nitrogen vapor phase
Derivation
The line was established in September 1988 from lymph node, a metastatic site.
Clinical Data
The patient received prior chemotherapy and radiation therapy.
The patient was a smoker. 52 pack years.

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
  1. 0.005 mg/ml Insulin
  2. 0.01 mg/ml Transferrin
  3. 30nM Sodium selenite (final conc.)
  4. 10 nM Hydrocortisone (final conc.)
  5. 10 nM beta-estradiol (final conc.)
  6. extra 2mM L-glutamine (for final conc. of 4.5 mM)
  7. 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.
  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 with 5% FBS and aspirate cells by gently pipetting.
  5. Transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to10 minutes.
  6. Discard supernatant and resuspend cells in fresh culture medium. Add appropriate aliquots of cell suspension to new culture vessels.
  7. Place culture vessels in incubators at 37°C.

Subcultivation Ratio: 1:3 to 1:5
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
Culture medium, 90%; additional fetal bovine serum, 5%; DMSO, 5%. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.
Culture Conditions
Temperature: 37°C

Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%

STR Profile
Amelogenin: X
CSF1PO: 10, 11
D13S317: None detected
D16S539: 12
D5S818: 10, 11
D7S820: 9, 11
THO1: 6, 9.3
TPOX: 8
vWA: 15, 20
Name of Depositor AF Gazdar, JD Minna
Deposited As Homo sapiens
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.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online.

Basic Documentation
Restrictions

The line is available with the following restrictions:

  1. This cell line was deposited at the ATCC by Dr. Adi F. Gazdar 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 National Cancer Institute (NCI).  For further information, please contact NCI’s Technology Transfer Center at NCI_TTC_Contact@mail.nih.gov or by phone at (240)-276-5514.

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.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online.