Bid KO SV40 MEF (ATCC® CRL-2911)

Organism: Mus musculus  /  Cell Type: fibroblast  /  Tissue: embryonic fibroblast  / 

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Organism Mus musculus
Tissue embryonic fibroblast
Cell Type fibroblast
Product Format frozen
Morphology fibroblast-like
Culture Properties adherent
Biosafety Level 2
Applications This cell line is useful to study molecular mechanism of cell apoptosis, BCL-2 family pathway, BID function, as well as TNF initiated cell death pathway.
Storage Conditions liquid nitrogen vapor phase
Images CRL-2911 Micrograph
Derivation Cells are immortalized MEFs which generated from BID knockout mice.
Tumorigenic no
Comments

Cells are immortalized MEFs (genotype: bid-/-), which were generated from BID knockout mice. 

BID is a pro-apoptotic BCL-2 family member. Cells display delayed apoptosis in response to TNF-alpha induced death due to a substantial reduction in the release of cytochrome c from the mitochondria and a reduction of the associated downstream effector caspase activity. 

This cell line is useful to study molecular mechanism of cell apoptosis, BCL-2 family pathway, BID function, as well as TNF initiated cell death pathway.

Complete Growth Medium The base medium for this cell line is ATCC-formulated IMDM Catalog No. 30-2005. To make the complete growth medium, add the following components to the base medium: 10 % Fetal Bovine Serum and 1x Non-essential amino acids.
Subculturing Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. 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.
  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 6.0 to 8.0 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 5 X 103 to 1 X 104 viable cells/cm2 is recommended.
  6. Incubate cultures at 37°C. At subculture, cell concentration is between 8 X 104 to 1.5 X 105 cells/cm2.
Subcultivation ratio: A subcultivation ratio of 1:8 to 1:20 is recommended.
Medium renewal: Every 2 to 3 days
Culture Conditions Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Name of Depositor Stanley Korsmeyer
References

Yin XM, et al. Bid-deficient mice are resistant to Fas-induced hepatocellular apoptosis. Nature 400 (6747) : 886-91, 1999 PubMed: 10476969

Cheng EH, et al. BCL-2, BCL-XL Sequester BH3 Domain-Only Molecules Preventing BAX- and BAK-Mediated Mitochondrial Apoptosis. Molecular Cell 8(3) : 705–711, 2001 PubMed: 11583631

Notice: Necessary PermitsPermits

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
Other Documentation
References

Yin XM, et al. Bid-deficient mice are resistant to Fas-induced hepatocellular apoptosis. Nature 400 (6747) : 886-91, 1999 PubMed: 10476969

Cheng EH, et al. BCL-2, BCL-XL Sequester BH3 Domain-Only Molecules Preventing BAX- and BAK-Mediated Mitochondrial Apoptosis. Molecular Cell 8(3) : 705–711, 2001 PubMed: 11583631