Cyt c-/- (ATCC® CRL-2613)

Organism: Mus musculus, mouse  /  Tissue: embryo  / 

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Organism Mus musculus, mouse
Tissue embryo
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Age 8-9 days gestation
Applications
The Cyt c-/- cell line provides a unique resource of mammalian cells lacking cytochrome c.
Derivation
The Cyt c -/- cell line was derived from a cytochrome c null mouse embryo at E8/E9 (8-9 days gestation) prior to embryonic death.
Comments
Murine embryos devoid of cytochrome c die in utero by midgestation. 
Cytochrome c released from mitochondria has been proposed to be an essential component of an apoptotic pathway responsive to DNA damage and other forms of cell stress.
The Cyt c -/- cells are devoid of cytochrome c and are resistant to various types of apoptotic stimuli.
They show reduced caspase-3 activation and are resistant to the proapoptotic effects of UV irradiation, serum withdrawal, and staurosporine. They demonstrate increased sensitivity to cell death signals triggered by TNFalpha.

Complete Growth Medium Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose and supplemented with: 25 mM HEPES, 0.1 mM nonessential amino acids, an additional 2 mM L-glutamine, 0.05 mM 2-mercaptoethanol, 0.05 mg/ml uridine, 500-1000 U/ml mouse leukemia inhibitory factor (LIF) and 20% fetal bovine serum
Subculturing
Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
  3. 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.
  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.
  6. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:5 is recommended
Medium Renewal: Every 3 days
Cryopreservation
Cell Culture Freezing Medium, 100% (GIBCO-BRL, Cat. No. 11101)
Culture Conditions
Temperature: 37°C
Name of Depositor RS Williams
References

Li K, et al. Cytochrome c deficiency causes embryonic lethality and attenuates stress-induced apoptosis. Cell 101: 389-399, 2000. PubMed: 10830166

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
References

Li K, et al. Cytochrome c deficiency causes embryonic lethality and attenuates stress-induced apoptosis. Cell 101: 389-399, 2000. PubMed: 10830166