Human Cell Lines-Cell Lines-InCellGene LLC.-CellLines,Biomedicine reasearch Product,Cell Biology,Cell Culture,293T Cell,Media

Organism	Homo sapiens, human
Tissue	mammary gland /breast
Cell Type	epithelial
Product Format	frozen
Morphology	epithelial
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.
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 Ref glucocorticoid receptor, positive Ref
Genes Expressed	luciferase. Ref 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).
Cellular Products	luciferase Ref(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