MDA-kb2 ATCC ® CRL-2713™ Homo sapiens mammary gland /breast

Permits and Restrictions	View Permits
Organism	Homo sapiens, human
Tissue	mammary gland /breast
Cell Type	epithelial
Product Format	frozen
Morphology	epithelial
Culture Properties	adherent
Biosafety Level	1
Age	48 years adult
Gender	female
Ethnicity	Caucasian
Applications	The cell line is an in vitro tool for research in endocrine activity of the human androgen receptor and glucocorticoid receptor. It can be used to screen chemicals for both androgen receptor and glucocorticoid receptor mediated activities.
Storage Conditions	liquid nitrogen vapor phase

Derivation	The MDA-kb2 cell line was derived from the breast cancer cell line, MDA-MB-453 (see ATCC HTB-131) by stable transfection with a mouse mammary tumor virus (MMTV) luciferase-neo reporter gene construct.
Clinical Data	48 years adult Caucasian female
Receptor Expression	androgen receptor RefWilson VS, et al. A novel cell line, MDA-kb2, that stably expresses an androgen- and glucocorticoid-responsive reporter for the detection of hormone receptor agonists and antagonists. Toxicol. Sci. 66: 69-81, 2002. PubMed: 11861974 glucocorticoid receptor, positive RefWilson VS, et al. A novel cell line, MDA-kb2, that stably expresses an androgen- and glucocorticoid-responsive reporter for the detection of hormone receptor agonists and antagonists. Toxicol. Sci. 66: 69-81, 2002. PubMed: 11861974
Genes Expressed	luciferase. RefWilson VS, et al. A novel cell line, MDA-kb2, that stably expresses an androgen- and glucocorticoid-responsive reporter for the detection of hormone receptor agonists and antagonists. Toxicol. Sci. 66: 69-81, 2002. PubMed: 11861974 The cell line expresses firefly luciferase under control of the MMTV promoter that contains response elements for both glucocorticoid receptors (GR) and androgen receptors (AR).
Comments	Since both glucocorticoid receptor and androgen receptor are present in the MDA-MB-453 cells, and both receptors can act through the MMTV promoter, compounds that act through either androgen receptor or glucocorticoid receptor activate the MMTV luciferase reporter. Androgen receptor agonists such as dihydrotestosterone, and glucocorticoid receptor agonists such as dexamethasone, corticosterone, and aldosterone induce luciferase expression. Responsiveness of the cell line was monitored over time and was stable for more than 80 passages.

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 10%. (Note: The L-15 medium formulation was devised for use in a free gas exchange with atmospheric air. A CO2 and air mixture is detrimental to cells when using this medium for cultivation)
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. Remove and discard culture medium. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.03% (w/v) EDTA solution to remove all traces of serum that contains trypsin inhibitor. 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. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C without CO₂. Subculture Ratio: 1:2 to 1:4 Medium Renewal: Every 2 to 3 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	Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase
Culture Conditions	Atmosphere: air, 100% Temperature: 37°C

STR Profile	Amelogenin: X CSF1PO: 10,12 D13S317: 12 D16S539: 9 D5S818: 11 D7S820: 10 THO1: 6 TPOX: 10 vWA: 17,18

Name of Depositor	VS Wilson
Passage History	Responsiveness of the cell line was monitored over time and was stable for more than 80 passages.
Year of Origin	November 25, 1998
References	Wilson VS, et al. A novel cell line, MDA-kb2, that stably expresses an androgen- and glucocorticoid-responsive reporter for the detection of hormone receptor agonists and antagonists. Toxicol. Sci. 66: 69-81, 2002. PubMed: 11861974 The MDA-kb2 cell line was derived from the breast cancer cell line, MDA-MB-453 (see ATCC HTB-131) by stable transfection with a mouse mammary tumor virus (MMTV) luciferase-neo reporter gene construct. The cell line expresses firefly luciferase under control of the MMTV promoter that contains response elements for both glucocorticoid receptors (GR) and androgen receptors (AR). MDA-kb2 may be used in an in vitro assay to screen androgen agonist and antagonists and to characterize its specificity and sensitivity to endocrine disrupting chemicals.

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 MSDS
References	Wilson VS, et al. A novel cell line, MDA-kb2, that stably expresses an androgen- and glucocorticoid-responsive reporter for the detection of hormone receptor agonists and antagonists. Toxicol. Sci. 66: 69-81, 2002. PubMed: 11861974 The MDA-kb2 cell line was derived from the breast cancer cell line, MDA-MB-453 (see ATCC HTB-131) by stable transfection with a mouse mammary tumor virus (MMTV) luciferase-neo reporter gene construct. The cell line expresses firefly luciferase under control of the MMTV promoter that contains response elements for both glucocorticoid receptors (GR) and androgen receptors (AR). MDA-kb2 may be used in an in vitro assay to screen androgen agonist and antagonists and to characterize its specificity and sensitivity to endocrine disrupting chemicals.

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

MSDS

References

Wilson VS, et al. A novel cell line, MDA-kb2, that stably expresses an androgen- and glucocorticoid-responsive reporter for the detection of hormone receptor agonists and antagonists. Toxicol. Sci. 66: 69-81, 2002. PubMed: 11861974

The MDA-kb2 cell line was derived from the breast cancer cell line, MDA-MB-453 (see ATCC HTB-131) by stable transfection with a mouse mammary tumor virus (MMTV) luciferase-neo reporter gene construct.

The cell line expresses firefly luciferase under control of the MMTV promoter that contains response elements for both glucocorticoid receptors (GR) and androgen receptors (AR).

MDA-kb2 may be used in an in vitro assay to screen androgen agonist and antagonists and to characterize its specificity and sensitivity to endocrine disrupting chemicals.

MDA-kb2 (ATCC® CRL-2713™)

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MDA-kb2 (ATCC^® CRL-2713^™)