Mfn1/Mfn2-null MEFs (ATCC® CRL-2994)

Organism: Mus musculus, mouse  /  Cell Type: fibroblast  /  Tissue: embryo/embryo fibroblast  / 

Permits and Restrictions

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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]
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 Mfn1 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.
  1. Remove and discard culture medium.
  2. 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.
  3. 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. 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

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
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