運(yùn)輸方式:凍存運(yùn)輸
數(shù)量:大量
器官來源:胚胎
年限:8-9 days gestation
生長狀態(tài):貼壁生長
細(xì)胞形態(tài):上皮樣
是否是腫瘤細(xì)胞:0
物種來源:小鼠
ATCC Number:CRL-2613? Designations: Cyt c-/-
Depositors: ?RS Williams
Biosafety Level:1
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:adherent
Organism:
Mus musculus deposited as mouse
Morphology:epithelial
Source:
Organ: embryo
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Age: 8-9 days gestation
Comments:Murine embryos devoid of cytochrome c die in utero by midgestation. 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. [51585 ]
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. [51585 ]
The Cyt c -/- cells are devoid of cytochrome c and are resistant to various types of apoptotic stimuli. [51585 ]
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. [51585 ]
The Cyt c-/- cell line provides a unique resource of mammalian cells lacking cytochrome c.
Propagation:
ATCC 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
Temperature: 37.0°C
Subculturing:
Protocol:
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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.
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Incubate cultures at 37?C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:5 is recommended
Medium Renewal: Every 3 days
Preservation: Cell Culture Freezing Medium, 100% (GIBCO-BRL, Cat. No. 11101)
Related Products:Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002
recommended serum:ATCC 30-2020
References: 51585: Li K, et al. Cytochrome c deficiency causes embryonic lethality and attenuates stress-induced apoptosis. Cell 101: 389-399, 2000. PubMed: 10830166