| 產(chǎn)品名稱 |
Cyt c-/- |
| 商品貨號(hào) |
B164309 |
| Organism |
Mus musculus, mouse |
| Tissue |
embryo |
| 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 |
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.
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| 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
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| 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.
- Remove and discard culture medium.
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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.
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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.
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Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
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Add appropriate aliquots of the cell suspension to new culture vessels.
- 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 |
| Deposited As |
mouse |
| References |
Li K, et al. Cytochrome c deficiency causes embryonic lethality and attenuates stress-induced apoptosis. Cell 101: 389-399, 2000. PubMed: 10830166
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