生长状态:混合型生长
是否是肿瘤细胞:0
物种来源:仓鼠
运输方式:冻存运输
器官来源:卵巢
细胞形态:上皮样
ATCC Number:CRL-1861?
数量:大量
规格:0.5ml Designations: EM9 (DNA repair mutant of CHO)
Depositors: ?LH Thompson
Biosafety Level:1
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:mixed, adherent and suspension
Organism:
Cricetulus griseus
Morphology:epithelial-like
Source:
Organ: ovary
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Gender: female
Comments:This line is a derivative of the CHO-K1 cell line (see ATCC CCL-61 ).
EM9 is a repair deficient mutant derived from AA8 (see ATCC CRL-1859 ).
The line was selected for enhanced sensitivity to ethylmethanesulfonate (EMS).
The line is defective in single strand break repair, has a 10 fold higher baseline frequency of sister chromatid exchange relative to AA8 and a 2 fold greater sensitivity to killing by X-rays.
This defect is corrected by the human XRCC1 gene.
Propagation:
ATCC complete growth medium: Alpha minimum essential medium without ribonucleosides and deoxyribonucleosides, 90%; fetal bovine serum, 10%
Temperature: 37.0°C
Subculturing:
Protocol:
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To subculture attaced cells, remove culture medium.
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The suspended cells are viable and can be used to start new cultures.
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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 1:4 to 1:12 is recommended
Medium Renewal: 2 to 3 times per week
Preservation:
Freeze medium: Complete growth medium 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Related Products:recommended serum:ATCC 30-2020
References: 1771: Thompson LH, et al. A CHO-cell strain having hypersensitivity to mutagens, a defect in DNA strand-break repair, and an extraordinary baseline frequency of sister-chromatid exchange. Mutat. Res. 95: 427-440, 1982. PubMed: 6889677
58396: Thompson LH, et al. A screening method for isolating DNA repair-deficient mutants of CHO cells. Somatic Cell Genet. 6: 391-405, 1980. PubMed: 7404270