OPA1-null MEFs細(xì)胞

年限： embryo, 10.5-days gestation

運(yùn)輸方式： 凍存運(yùn)輸

數(shù)量： 大量

組織來源： embryo fibroblast

細(xì)胞形態(tài)： 成纖維樣

器官來源： 胚胎

生長(zhǎng)狀態(tài)： 貼壁生長(zhǎng)

是否是腫瘤細(xì)胞： 0

物種來源： 小鼠

ATCC Number： CRL-2995?

Designations: OPA1-null MEFs

Depositors: D. Chan

Biosafety Level: 2

OPA1-null MEFs細(xì)胞Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Mus musculus

Morphology: fibroblast-like

Source: Organ: embryo

Tissue: embryo fibroblast

Breed: 129/SvEv x C57BL/6

Donor Organism Characteristics: knockout

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: 2005

Age: embryo, 10.5-days gestation

Comments: OPA1-null MEFs細(xì)胞Mitochondrial fusion plays an important role in controlling the morphology and function of mitochondria. In mammalian cells, the dynamin-related GTPase OPA1 is essential for mitochondrial fusion. OPA1 is associated with the inner membranes and protects the cells from apoptosis by regulating inner membrane dynamics. Mutation of OPA1 leads to degeneration of the retinal ganglion cells that causes dominant optic atrophy. OPA1-null cells have been developed as a cellular system to study how individual OPA1 splice forms contribute to mitochondrial fusion.

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: fetal bovine serum to a final concentration of 10%.

Temperature: 37.0°C

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

Subculturing: Protocol: Volumes used in this protocol are for 75 sq. cm. 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.

Incubate cultures at 37.0°C.

OPA1-null MEFs細(xì)胞Subcultivation ratio: A subcultivation ratio of 1:5 to 1:20 is recommended.

Medium renewal: Every 2 to 3 days

Preservation: Freeze medium: complete growth medium, 90%; DMSO, 10%

liquid nitrogen vapor phase

Related Products: Recommended medium (without the additional serum described under ATCC Medium): ATCC 30-2002

Recommended serum: ATCC 30-2020

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

Cell culture tested DMSO: ATCC 4-X

Erythrosin B vital stain solution: ATCC 30-2404

Related cell line: ATCC CRL-2991

Related cell line: ATCC CRL-2992

Related cell line: ATCC CRL-2993

Related cell line: ATCC CRL-2994

References: 16173333: Chen H, et al. Mitofusins Mfn1 and Mfn2 coordinately regulate mitochondrial fusion and are essential for embryonic development. J. Cell Biol. 160: 189-200, 2003. PubMed: 12527753

16173335: Song Z, et al. OPA1 processing controls mitochondrial fusion and is regulated by mRNA splicing, membrane potential, and Yme1L. J. Cell Biol. 178(5): 749-755, 2007. PUbMed: 17709429

16173402: Cipolat S, et. al. OPA1-null MEFs細(xì)胞OPA1 requires mitofusin 1 to promote mitochondrial fusion. Proc. Natl. Acad. Sci. USA. 101(45): 15927-15932, 2004. PubMed: 15509649