組織來源:embryo
生長狀態(tài):貼壁生長
器官來源:胚胎
運(yùn)輸方式:凍存運(yùn)輸
細(xì)胞形態(tài):成纖維樣
是否是腫瘤細(xì)胞:0
物種來源:小鼠
年限:embryo, 13.5 days gestation
ATCC Number:CRL-2979?
數(shù)量:大量
規(guī)格:0.1ml Designations: CHOP-KO-DR
Depositors: ?D Ron and H Harding
Biosafety Level:2
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:adherent
Organism:
Mus musculus
Morphology:fibroblast-like
Source:
Organ: embryo
Tissue: embryo
Cellular Products:C/EBP homologous protein (CHOP)/growth arrest and DNA damage-inducible gene 153 (GADD153), not expressed
Permits/Forms:In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Isolation: Isolation date: 1998
Applications:CHOP is a major regulator of the endoplasmic reticulum UPR and as such, these CHOP-KO cells are useful for studying the mechanisms of UPR and integrated stress response.
DR-Wildtype cells, CRL-2977, are available for use as a control.
These cells lack the transcription factor CHOP, a key mediator of cell death in the unfolded protein response (UPR).
Age: embryo, 13.5 days gestation
Comments:The mouse embryonic fibroblast (MEF) cell line, CHOP-KO-DR (CRL-2979), was established from a 13.5 day-old CHOP-/- mouse embryo by SV-40 immortalization. These cells lack the transcription factor CHOP, a key mediator of cell death in the unfolded protein response (UPR). CHOP is a major regulator of the endoplasmic reticulum UPR and as such, these CHOP-KO cells are useful for studying the mechanisms of UPR and integrated stress response. DR-Wildtype cells, CRL-2977, are available for use as a control.
Propagation:
ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium:
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O.1 mM Non-Essential Amino Acids (NEAA)
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0.05mM 2-Mercaptoethanol
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fetal bovine serum to a final concentration of 10%
Temperature: 37.0°C
Atmosphere: air, 95%; carbon dioxide (CO
2 ), 5%
Subculturing:
Protocol: Volumes used in this protocol are for 75 cm
2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
-
Remove and discard culture medium.
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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.
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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 37.0°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. An inoculum of 2 X 103 to 3 X 103 viable cells/cm2 is recommended.
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Incubate cultures at 37.0°C.
Subcultivation ratio: A subcultivation ratio of 1:10 to 1:30 twice weekly is recommended.
Medium renewal: every 2 to 3 days
Preservation:
Freeze medium: fetal bovine serum, 90%; DMSO, 10%
Storage temperature: liquid nitrogen vapor phase
Related Products:Recommended medium (without the additional serum described under ATCC Medium): ATCC 30-2002
Recommended serum: ATCC 30-2020
MEM Non-Essential Amino Acid Solution, 100x: ATCC 30-2116
Cell culture tested DMSO: ATCC 4-X
0.25% (w/v) Trypsin - 0.53mM EDTA in Hank's BSS (w/o Ca++, Mg ++): ATCC 30-2101
Phosphate-buffered saline: ATCC 30-2200
References: 16173432: Zinszner H, et al. CHOP is implicated in programmed cell death in response to impaired function of the endoplasmic reticulum. Genes Dev. 12(7): 982-995, 1998. PubMed: 9531536