T47D-KBluc (ATCC® CRL-2865)

Organism: Homo sapiens, human  /  Cell Type: epithelial transfected with reporter plasmid  /  Tissue: mammary gland; breast/duct; derived from metastatic site: pleural effusion  /  Disease: ductal carcinoma

Permits and Restrictions

View Restrictions

Organism Homo sapiens, human
Tissue mammary gland; breast/duct; derived from metastatic site: pleural effusion
Cell Type epithelial transfected with reporter plasmid
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease ductal carcinoma
Age 54 years
Gender female
Applications
They can be used to screen chemicals for estrogenic or anti-estrogenic activity.
Storage Conditions liquid nitrogen vapor phase
Clinical Data
female
HeLa Markers N
Comments

The parent cell line was transfected with pGL2.TATA.Inr.luc.neo which contains three estrogen responsive elements upstream of a luc reporter gene. 

The cells were selected for responsiveness to 17-beta-estradiol. They can be used to screen chemicals for estrogenic or anti-estrogenic activity.

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: 0.2 Units/ml bovine insulin; fetal bovine serum to a final concentration of 10%.
Subculturing Volumes used in this protocol are for 75cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Note: Subculture cells before or upon reaching confluence. Do not allow cells to become super-confluent.

  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with either Hanks' Balanced Salt Solution (HBSS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
  3. 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°C to facilitate dispersal.
  4. Add 10 to 15 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 2 x 104 to 4 x 104 viable cells/cm2 is recommended.
  6. Incubate cultures at 37C. We recommend that you maintain cultures at a cell concentration between 3 x 104 and 2 x 105 cells/cm2.

Subcultivation Ratio: 1:2 to 1:3 is recommended
Medium Renewal: every 2 to 3 days
Cryopreservation
Freeze medium: culture medium, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Duration: The depositor states that supplementing the growth medium with insulin is optional.
STR Profile
Amelogenin: X
CSF1PO: 11,13
D13S317: 12
D16S539: 10
D5S818: 12
D7S820: 11
THO1: 6
TPOX: 11
vWA: 14
Population Doubling Time 36
Name of Depositor VS Wilson
Year of Origin January 4, 2000
References

Wilson VS, et al. Development and characterization of a cell line that stably expresses an estrogen-responsive luciferase reporter for the detection of estrogen receptor agonist and antagonists. Toxicol. Sci. 81: 69-77, 2004. PubMed: 15166400

Basic Documentation
Restrictions

This product's use is governed by a Label License. For information on purchasing a license to use this product for purposes other than those permitted in the Label License, please contact Promega.

References

Wilson VS, et al. Development and characterization of a cell line that stably expresses an estrogen-responsive luciferase reporter for the detection of estrogen receptor agonist and antagonists. Toxicol. Sci. 81: 69-77, 2004. PubMed: 15166400