UACC-3133 ATCC ® CRL-2988™ Homo sapiens breast infiltrating

Permits and Restrictions	View Permits
Organism	Homo sapiens, human
Tissue	breast
Cell Type	epithelial-like
Product Format	frozen
Morphology	epithelial-like
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	infiltrating lobular carcinoma of breast
Age	52 years old
Gender	female
Ethnicity	Caucasian
Storage Conditions	liquid nitrogen vapor phase

Karyotype	range=34-155; modal number=40-42
Derivation	This human breast cancer cell line, UACC-3133, was derived from a 52 year-old female with poorly differentiated adenocarcinoma, which is, consistent with infiltrating lobular carcinoma of breast metastatic to the pleural fluid. Prior to surgery, the patient received no chemotherapy.
Receptor Expression	neu (ErbB2; HER2), expressed estrogen receptor (ER), low expression epidermal growth factor receptor (EGFR), not expressed progesterone receptor (PR), not expressed
Comments	These cells have very low expression level of estrogen receptors, and are negative for progesterone receptors. They are negative for the tumor suppressor gene MASPIN and cell adhesion molecule DSC3. This cell line has unique expression profile of bone morphogenetic proteins (BMPs) which belong to the TGF-beta superfamily. The cells have high level of BMP3, and lack of BMP2 expression.

Complete Growth Medium	The base medium for this cell line is ATCC-formulated Leibovitz's L-15 Medium, Catalog No. 30-2008. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 5% 0.01 mg/ml transferrin (final conc.) 0.01 mg/ml insulin (final conc.) 5 µg/ml (55 U/ml) catalase (final conc.) 3.6 µg/ml (0.01 mM) hydrocortisone (final conc.) extra 2 mM glutamine
Subculturing	Note: This cell line grows slowly Volumes used in this protocol are for 75 cm² flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Remove and discard culture medium. 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 to remove all traces of serum which contains trypsin inhibitor. Add 1.0 to 2.0 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 37.0°C to facilitate dispersal. 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 xg for 5 to 10 minutes. Discard supernatant. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 1.5 X 10⁴ to 2.5 x 10⁴ viable cells/cm² is recommended. Incubate cultures at 37.0°C. Subculture when the cell concentration is between 3 X 10⁴ to 5 x 10⁴ cells/cm². Subcultivation ratio: A subcultivation ratio of 1:2 is recommended. Medium renewal: every 3 to 4 days
Cryopreservation	Freeze medium: complete growth medium, 80%; FBS, 10%; DMSO, 10% Storage temperature: liquid nitrogen vapor phase
Culture Conditions	Temperature: 37°C Atmosphere: air, 100% Growth Conditions: This cell line grows slowly.

STR Profile	D5S818: 9, 11 D13S317: 13 D7S820: 8, 10 D16S539: 11 vWA: 16, 17 TH01: 9, 3 Amelogenin: X TPOX: 8 CSF1PO: 12,13

Name of Depositor	K Brown
References	Domann FE, et al. Epigenetic silencing of maspin gene expression in human breast cancers. Int. J. Cancer. 85(6): 805-810, 2000 PubMed: 10709100 Kauraniemi P, et al. New amplified and highly expressed genes discovered in the ERBB2 amplicon in breast cancer by cDNA microarrays. Cancer Res. 61(22): 8235-8240, 2001 PubMed: 117194555 Ortiz RM, et al. Aberrant alternative exon use and increased copy number of human metalloprotease-disintegrin ADAM15 gene in breast cancer cells. Genes Chromosomes Cancer. 41(4): 366-378, 2004. PubMed: 15384173 Futscher BW, et al. Chapter 6: Epigenetic Dysregulation of Maspin (SerpinB5) in Cancer Invasion and Metastasis. DNA Methylation, Epigenetics and Metastasis. Cancer Metastasis - Biology and Treatment, 7: 133-155, 2005. Oshiro MM, et al. Epigenetic silencing of DSC3 is a common event in human breast cancer. Breast Cancer Res. 7(5): R669-R680, 2005. PubMed: 16168112 Alarmo EL, et al. Bone morphogenetic protein 7 is widely overexpressed in primary breast cancer. Genes Chromosomes Cancer. 45(4): 411-419, 2006. PubMed: 16419056 Futscher BW, et al. Chapter 7: Epigenetic Regulation of Genes That Affect Tumor Cell Adhesion. Cell Adhesion and Cytoskeletal Molecules in Metastasis. Cancer Metastasis - Biology and Treatment, 9: 123-140, 2006. Alarmo EL, et al. A comprehensive expression survey of bone morphogenetic proteins in breast cancer highlights the importance of BMP4 and BMP7. Breast Cancer Res. Treat. 103(2): 239-246, 2007. PubMed: 17004110 Parssinen J, et al. PPM1D silencing by RNA interference inhibits proliferation and induces apoptosis in breast cancer cell lines with wild-type p53. Cancer Genet. Cytogenet. 182(1): 33-39, 2008. PubMed: 18328948 Robey IF, et al. Regulation of the warburg effect in early-passage breast cancer cells. Neoplasia 10(8): 745-756, 2008. PubMed: 18670636 Ketolainen JM, et al. Parallel inhibition of cell growth and induction of cell migration and invasion in breast cancer cells by bone morphogenetic protein 4. Breast Cancer Res. Treat. 124(2): 377-386, 2010. PubMed: 20182795

Name of Depositor

K Brown

References

Domann FE, et al. Epigenetic silencing of maspin gene expression in human breast cancers. Int. J. Cancer. 85(6): 805-810, 2000 PubMed: 10709100

Kauraniemi P, et al. New amplified and highly expressed genes discovered in the ERBB2 amplicon in breast cancer by cDNA microarrays. Cancer Res. 61(22): 8235-8240, 2001 PubMed: 117194555

Ortiz RM, et al. Aberrant alternative exon use and increased copy number of human metalloprotease-disintegrin ADAM15 gene in breast cancer cells. Genes Chromosomes Cancer. 41(4): 366-378, 2004. PubMed: 15384173

Futscher BW, et al. Chapter 6: Epigenetic Dysregulation of Maspin (SerpinB5) in Cancer Invasion and Metastasis. DNA Methylation, Epigenetics and Metastasis. Cancer Metastasis - Biology and Treatment, 7: 133-155, 2005.

Oshiro MM, et al. Epigenetic silencing of DSC3 is a common event in human breast cancer. Breast Cancer Res. 7(5): R669-R680, 2005. PubMed: 16168112

Alarmo EL, et al. Bone morphogenetic protein 7 is widely overexpressed in primary breast cancer. Genes Chromosomes Cancer. 45(4): 411-419, 2006. PubMed: 16419056

Futscher BW, et al. Chapter 7: Epigenetic Regulation of Genes That Affect Tumor Cell Adhesion. Cell Adhesion and Cytoskeletal Molecules in Metastasis. Cancer Metastasis - Biology and Treatment, 9: 123-140, 2006.

Alarmo EL, et al. A comprehensive expression survey of bone morphogenetic proteins in breast cancer highlights the importance of BMP4 and BMP7. Breast Cancer Res. Treat. 103(2): 239-246, 2007. PubMed: 17004110

Parssinen J, et al. PPM1D silencing by RNA interference inhibits proliferation and induces apoptosis in breast cancer cell lines with wild-type p53. Cancer Genet. Cytogenet. 182(1): 33-39, 2008. PubMed: 18328948

Robey IF, et al. Regulation of the warburg effect in early-passage breast cancer cells. Neoplasia 10(8): 745-756, 2008. PubMed: 18670636

Ketolainen JM, et al. Parallel inhibition of cell growth and induction of cell migration and invasion in breast cancer cells by bone morphogenetic protein 4. Breast Cancer Res. Treat. 124(2): 377-386, 2010. PubMed: 20182795

Permits	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	Product Sheet Certificate of Analysis SDS
References	Domann FE, et al. Epigenetic silencing of maspin gene expression in human breast cancers. Int. J. Cancer. 85(6): 805-810, 2000 PubMed: 10709100 Kauraniemi P, et al. New amplified and highly expressed genes discovered in the ERBB2 amplicon in breast cancer by cDNA microarrays. Cancer Res. 61(22): 8235-8240, 2001 PubMed: 117194555 Ortiz RM, et al. Aberrant alternative exon use and increased copy number of human metalloprotease-disintegrin ADAM15 gene in breast cancer cells. Genes Chromosomes Cancer. 41(4): 366-378, 2004. PubMed: 15384173 Futscher BW, et al. Chapter 6: Epigenetic Dysregulation of Maspin (SerpinB5) in Cancer Invasion and Metastasis. DNA Methylation, Epigenetics and Metastasis. Cancer Metastasis - Biology and Treatment, 7: 133-155, 2005. Oshiro MM, et al. Epigenetic silencing of DSC3 is a common event in human breast cancer. Breast Cancer Res. 7(5): R669-R680, 2005. PubMed: 16168112 Alarmo EL, et al. Bone morphogenetic protein 7 is widely overexpressed in primary breast cancer. Genes Chromosomes Cancer. 45(4): 411-419, 2006. PubMed: 16419056 Futscher BW, et al. Chapter 7: Epigenetic Regulation of Genes That Affect Tumor Cell Adhesion. Cell Adhesion and Cytoskeletal Molecules in Metastasis. Cancer Metastasis - Biology and Treatment, 9: 123-140, 2006. Alarmo EL, et al. A comprehensive expression survey of bone morphogenetic proteins in breast cancer highlights the importance of BMP4 and BMP7. Breast Cancer Res. Treat. 103(2): 239-246, 2007. PubMed: 17004110 Parssinen J, et al. PPM1D silencing by RNA interference inhibits proliferation and induces apoptosis in breast cancer cell lines with wild-type p53. Cancer Genet. Cytogenet. 182(1): 33-39, 2008. PubMed: 18328948 Robey IF, et al. Regulation of the warburg effect in early-passage breast cancer cells. Neoplasia 10(8): 745-756, 2008. PubMed: 18670636 Ketolainen JM, et al. Parallel inhibition of cell growth and induction of cell migration and invasion in breast cancer cells by bone morphogenetic protein 4. Breast Cancer Res. Treat. 124(2): 377-386, 2010. PubMed: 20182795

Permits

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

Product Sheet

Certificate of Analysis

SDS