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Mfn1/Mfn2-null MEFs

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貨期:

編號:B161974

品牌:Mingzhoubio

標準菌株

定量菌液

DNA

RNA

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凍干粉

斜面

甘油

平板

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產品名稱

Mfn1/Mfn2-null MEFs

商品貨號

B161974

Organism

Mus musculus, mouse

Tissue

embryo/embryo fibroblast

Cell Type

fibroblast

Product Format

frozen

Morphology

fibroblast-like

Culture Properties

adherent

Biosafety Level

2 [Cells contain SV40 viral DNA sequences]

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Age

embryo, 10.5-days gestation

Applications

These cell lines are useful in studying mitochondrial biology and the role of mitochondrial fusion in cell physiology.

Storage Conditions

liquid nitrogen vapor phase

Images

Derivation

MEFs were derived from embryos 10.5 days gestation. Embryos were mechanically dispersed by repeated passage through a P1000 pipette tip and plated with MEF medium (DME, 10% FCS, 1X nonessential amino acids, 1 mM L-glutamine (Invitrogen/Gibco)). Cells with deletions of Mfn1 (ATCC CRL-2992) or Mfn2 (ATCC CRL-2993) or both Mfn1/Mfn2-null MEFs (ATCC CRL-2994) were subsequently immortalized by transduction with SV-40 T antigen.

Clinical Data

Donor Organism Characteristics: knockout

Comments

Both Mfn1 and Mfn2 are essential for mitochondrial fusion which is essential for embryonic development.

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%.

Subculturing

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°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°C.

Subcultivation ratio: A subcultivation ratio of 1:5 to 1:20 is recommended

Medium renewal: Every 2 to 3 days

Cryopreservation

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

liquid nitrogen vapor phase

Culture Conditions

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

Temperature: 37°C

Name of Depositor

D Chan

Year of Origin

2003

References

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

Koshiba T, et al. Structural basis of mitochondrial tethering by mitofusin complexes. Science 305(5685): 858-862, 2004. PubMed: 15297672

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