AU565 [AU-565] (ATCC® CRL-2351)

Organism: Homo sapiens, human  /  Tissue: mammary gland/breast; derived from metastitic site: malignant pleural effusion  /  Disease: adenocarcinoma

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Organism Homo sapiens, human
Tissue
mammary gland/breast; derived from metastitic site: malignant pleural effusion
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease adenocarcinoma
Age 43 years
Gender female
Ethnicity Caucasian, White
Applications
The cells are useful as models of cellular growth to study the differentiation of breast cancer cells.
Storage Conditions liquid nitrogen vapor phase
Images
Derivation
The AU565 cell line was derived at the Naval Biosciences Laboratory, Oakland, CA from a pleural effusion of a patient with breast carcinoma. This cell line was established from the same patient as SK-BR-3 (ATCC HTB-30).
Clinical Data
43 years
Caucasian, White
female
Receptor Expression
epidermal growth factor (EGF)
Oncogene her2/neu + (overexpressed); her3 +; her4 +; p53 +
Comments

The patient, a White, Caucasian female, age 43, blood type A+, had been treated with radiation, steroids, cytoxan and 5-fluorouracil.

The AU565 cell line amplifies and overexpresses the HER-2/neu oncogene; it expresses the HER-3, HER-4 and p53 oncogenes.

Treating the cells with mycophenolic acid (MPA), phorbol 12-myristate 13-acetate (PMA), retinoic acid (RA) or the TA-1 monoclonal antibody to the extracellular domain of the HER-2/neu protein can induce cell differentiation.

Treatment of AU565 cells with Neu differentiation factor (NDF) also induces mature phenotype, which is characterized by the morphological appearance of the cells.

Untreated AU565 cells have compact nuclei with thin cytoplasm while treated cells display a morphology associated with a differentiated phenotype such as flat large nuclei surrounded by a sizable cytoplasm.

A culture submitted to the ATCC in April of 1995 was found to be contaminated with mycoplasma and progeny were cured by a 21-day treatment with BM Cycline.

The cells were assayed for mycoplasma, by the Hoechst stain, PCR and the standard culture test, after a six-week period following treatment. All tests were negative.

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%.


Phenol Red is a structural mimic for estrogen. Therefore the estrogenic activity of phenol red should be considered in any studies that utilize estrogen-responsive cells in culture (RefBerthois Y, et al. Phenol red in tissue culture media is a weak estrogen: implications concerning the study of estrogen-responsive cells in culture. Proc. Natl. Acad. Sci. USA 83: 2496–2500, 1986. PubMed: 3458212). The Phenol Red-free version of RPMI 1640, is ATCC catalog No. 30-2602.

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.
Note: Do NOT allow cells to become confluent.  Subculture at 50-60% confluency.  The majority of cells should be round and undifferentiated.  If cells overgrow, they can be brought back to an undifferentiated condition by maintaining them under sparse conditions for a couple of passages. 

  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum that 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

Subculture Ratio: 1:4 to 1:6
Medium Renewal: Every 3 to 5 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
culture medium 95%; DMSO, 5%
Culture Conditions
Temperature: 37°C
STR Profile
Amelogenin: X
CSF1PO: 12
D13S317: 11,12
D16S539: 9
D5S818: 9,12
D7S820: 9,12
THO1: 8,9
TPOX: 8,11
vWA: 17
Name of Depositor SS Bacus
References

Bacus SS, et al. Differentiation of cultured human breast cancer cells (AU-565 and MCF-7) associated with loss of cell surface HER-2/neu antigen. Mol. Carcinog. 3: 350-362, 1990. PubMed: 1980588

Bacus SS, et al. Tumor-inhibitory monoclonal antibodies to the HER-2/Neu receptor induce differentiation of human breast cancer cells. Cancer Res. 52: 2580-2589, 1992. PubMed: 1373672

Bacus SS, et al. Neu differentiation factor (heregulin) induces expression of intercellular adhesion molecule 1: implications for mammary tumors. Cancer Res. 53: 5251-5261, 1993. PubMed: 8106145

Wen D, et al. Neu differentiation factor: a transmembrane glycoprotein containing an EGF domain and an immunoglobulin homology unit. Cell 69: 559-572, 1992. PubMed: 1349853

Bacus SS, et al. A ligand for the erbB-2 oncogene product (gp30) induces differentiation of human breast cancer cells. Cell Growth Differ. 3: 401-411, 1992. PubMed: 1358180

Peles E, et al. Isolation of the neu/HER-2 stimulatory ligand: a 44 kd glycoprotein that induces differentiation of mammary tumor cells. Cell 69: 205-216, 1992. PubMed: 1348215

Lessor T, et al. Regulation of heregulin beta1-induced differentiation in a human breast carcinoma cell line by the extracellular-regulated kinase (ERK) pathway. J. Cell. Biochem. 70: 587-595, 1998. PubMed: 9712155

Yoo JY, et al. Inhibition of cell proliferation by 17beta-estradiol and heregulin beta1 in estrogen receptor negative human breast carcinoma cell lines. Breast Cancer Res. Treat. 51: 71-81, 1998. PubMed: 9877030

Yoo JY, Hamburger AW. Changes in heregulin beta1 (HRGbeta1) signaling after inhibition of ErbB-2 expression in a human breast cancer cell line. Mol. Cell. Endocrinol. 138: 163-171, 1998. PubMed: 9685225

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
Other Documentation
References

Bacus SS, et al. Differentiation of cultured human breast cancer cells (AU-565 and MCF-7) associated with loss of cell surface HER-2/neu antigen. Mol. Carcinog. 3: 350-362, 1990. PubMed: 1980588

Bacus SS, et al. Tumor-inhibitory monoclonal antibodies to the HER-2/Neu receptor induce differentiation of human breast cancer cells. Cancer Res. 52: 2580-2589, 1992. PubMed: 1373672

Bacus SS, et al. Neu differentiation factor (heregulin) induces expression of intercellular adhesion molecule 1: implications for mammary tumors. Cancer Res. 53: 5251-5261, 1993. PubMed: 8106145

Wen D, et al. Neu differentiation factor: a transmembrane glycoprotein containing an EGF domain and an immunoglobulin homology unit. Cell 69: 559-572, 1992. PubMed: 1349853

Bacus SS, et al. A ligand for the erbB-2 oncogene product (gp30) induces differentiation of human breast cancer cells. Cell Growth Differ. 3: 401-411, 1992. PubMed: 1358180

Peles E, et al. Isolation of the neu/HER-2 stimulatory ligand: a 44 kd glycoprotein that induces differentiation of mammary tumor cells. Cell 69: 205-216, 1992. PubMed: 1348215

Lessor T, et al. Regulation of heregulin beta1-induced differentiation in a human breast carcinoma cell line by the extracellular-regulated kinase (ERK) pathway. J. Cell. Biochem. 70: 587-595, 1998. PubMed: 9712155

Yoo JY, et al. Inhibition of cell proliferation by 17beta-estradiol and heregulin beta1 in estrogen receptor negative human breast carcinoma cell lines. Breast Cancer Res. Treat. 51: 71-81, 1998. PubMed: 9877030

Yoo JY, Hamburger AW. Changes in heregulin beta1 (HRGbeta1) signaling after inhibition of ErbB-2 expression in a human breast cancer cell line. Mol. Cell. Endocrinol. 138: 163-171, 1998. PubMed: 9685225