GCN2-KO-DR細胞

運輸方式： 凍存運輸

器官來源： 胚胎

ATCC Number： CRL-2978?

是否是腫瘤細胞： 0

物種來源： 小鼠

細胞形態： 成纖維樣

生長狀態： 貼壁生長

年限： embryo, 13.5 days gestation

組織來源： embryo

數量： 大量

GCN2-KO-DR細胞Designations: GCN2-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: Breed: W4/129S6

Organ: embryo

Tissue: embryo

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. GCN2-KO-DR細胞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: 2000

Applications: GCN2 is a major regulator of the endoplasmic reticulum unfolded protein response (UPR) and as such, these KO cells are a useful tool for studying UPR. DR-Wildtype cells, CRL-2979, are available for use as a control.

The cells lack the nutritional stress-linked kinase GCN2, which phosphorylates eIF2a (alpha subunit of eukaryotic initiation factor 2) and complements PERK. GCN2-/- cells have been used to assess the role of eIF2a phosphorylation in a variety of pathophysiological conditions.

Age: embryo, 13.5 days gestation

Comments: The mouse embryonic fibroblast (MEF) cell line, CRL-2978 (GCN2-KO-DR), was established from a 13.5 day-old GCN2-/- mouse embryo by SV-40 immortalization. The cells lack the nutritional stress-linked kinase GCN2, which phosphorylates eIF2a (alpha subunit of eukaryotic initiation factor 2) and complements PERK. GCN2-KO-DR細胞GCN2-/- cells have been used to assess the role of eIF2a phosphorylation in a variety of pathophysiological conditions. GCN2 is a major regulator of the endoplasmic reticulum unfolded protein response (UPR) and as such, these KO cells are a useful tool for studying UPR. DR-Wildtype cells, CRL-2979, 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:

O.1 mM Non-Essential Amino Acids (NEAA)

0.05mM 2-Mercaptoethanol

fetal bovine serum to a final concentration of 10%

Temperature: 37.0°C

Atmosphere: air, 95%; carbon dioxide (CO2), 5%

Subculturing: GCN2-KO-DR細胞Protocol: Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

Remove and discard culture medium.

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.

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.

Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.

Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 1 X 103 to 2 X 103 viable cells/cm2 is recommended.

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:GCN2-KO-DR細胞 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: 16173431: Harding HP, et al. Regulated translation initiation controls stress-induced gene expression in mammalian cells. Mol. Cell 6(5): 1099-1108, 2000. PubMed: 11106749