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MDA-kb2

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價格:

編號:B165119

品牌:Mingzhoubio

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DNA

RNA

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產品名稱

MDA-kb2

商品貨號

B165119

Organism

Homo sapiens, human

Tissue

mammary gland /breast

培養基

Cell Type

epithelial

Product Format

frozen

Morphology

epithelial

Culture Properties

adherent

Biosafety Level

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. 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).

Cellular Products

luciferase

Ref

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 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

Deposited As

human

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

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.

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