HCC4006 (ATCC® CRL-2871)

Organism: Homo sapiens, human  /  Cell Type: epithelial  /  Tissue: lung; derived from metastatic site: pleural effusion  /  Disease: adenocarcinoma

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Organism Homo sapiens, human
Tissue lung; derived from metastatic site: pleural effusion
Cell Type epithelial
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease adenocarcinoma
Age greater than 50 years adult
Gender male
Ethnicity Caucasian
Storage Conditions liquid nitrogen vapor phase
Clinical Data
50+ years
male
Caucasian
Comments
This lung adenocarcinoma has an acquired mutation in the EGFR tyrosine kinase domain (L747 - E749 deletion, A750P).
Complete Growth Medium The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. 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 75cm2 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 or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution or Dulbecco’s Phosphate Buffered Saline (D-PBS) to remove all traces of serum that contains trypsin inhibitor.
  3. Add 1.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until the 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°Cto facilitate dispersal.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. 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. An inoculum of 5 x 103 to 7 x 103 viable cells/cm2 is recommended.
  7. Place culture vessels in incubators at 37°C. Maintain cultures at a cell concentration between 1 x 104 and 3 x 104 cells/2.

Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:6 is recommended
Medium Renewal: Every 2 to 3 days
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
Population Doubling Time 41 hours
Name of Depositor JD Minna, AF Gazdar
Year of Origin April 8, 2004
References

Shigematsu H, et al. Somatic mutations of the HER2 kinase domain in lung adenocarcinomas. Cancer Res. 65: 1642-1646, 2005. PubMed: 15753357

Although isolated from a tumor that arose in a male patient, a Y chromosome marker (amelogenin) was not detected at ATCC.

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) 648-1888, Email TechnologyDevelopment@UTSouthwestern.edu, or Fax: (214) 951-0935.

References

Shigematsu H, et al. Somatic mutations of the HER2 kinase domain in lung adenocarcinomas. Cancer Res. 65: 1642-1646, 2005. PubMed: 15753357

Although isolated from a tumor that arose in a male patient, a Y chromosome marker (amelogenin) was not detected at ATCC.