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This cell line was initiated from an axillary lymph node on 6/7/94 and took 12.5 months to establish.

A B lymphoblastoid cell line initiated by Epstein-Barr virus (EBV) transformation of peripheral blood lymphocytes from this patient is available as HCC1007 BL (CRL-2319)

The cells are positive for expressions of Her2-neu and p53 oncogenes.

HCC1008 is positive for the epithelial cell specific marker, Epithelial Glycoprotein 2 (EGP2) and cytokeratin 19.

The cells are negative for expression of estrogen receptor (ER) and for expression of progesterone receptor (PR).

A B lymphoblastoid cell line initiated by Epstein-Barr virus (EBV) transformation of peripheral blood lymphocytes from this patient is available as HCC1007 BL (ATCC CRL-2319)

• 0.005 mg/ml Insulin
• 0.01 mg/ml Transferrin
• 30nM Sodium selenite (final conc.)
• 10 nM Hydrocortisone (final conc.)
• 10 nM beta-estradiol (final conc.)
• extra 2mM L-glutamine (for final conc. of 4.5 mM)
• 10% fetal bovine serum (final conc.)

1. Remove and discard culture medium.
2. Add 3.0 to 4.0 mL Cell Dissociation Buffer (GIBCO #13150-016) to cell layer and incubate at room temperature or 37C.
3. Observe cells under an inverted microscope until cell layer is dispersed (usually with 5 to 10 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. To remove Cell Dissociation Buffer, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes.
6. Discard supernatant and resuspend cell pellet in fresh complete growth medium. Aspirate cells with a small bore pipette. Add appropriate aliquots of cell suspension to new culture vessels.
7. Place culture vessels in incubators at 37°C.

Note: This cell line grows exceedingly slow. Cells grow in patches and cultures only become 50 to 60% confluent.

Note: Cells reattach slowly after subculture (about 50% attached and 50% in suspension). Allow flasks to remain undisturbed for at least a week for cultures to become reestablished. It can take as along as three to four weeks before cultures can be subcultured again.

Subcultivation Ratio: A subcultivation ratio of 1:2 is recommended
Medium Renewal: Every 3 to 4 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.

Gazdar AF, et al. Characterization of paired tumor and non-tumor cell lines established from patients with breast cancer. Int. J. Cancer 78: 766-774, 1998. PubMed: 9833771

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.