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生長狀態(tài)：貼壁生長
器官來源：胚胎
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
ATCC Number：CRL-2993?
運輸方式：凍存運輸
數(shù)量：大量
組織來源：embryo fibroblast
年限：embryo, 10.5-days gestation
規(guī)格：0.2ml Designations: Mfn2-null MEFs
Depositors: ?D. Chan
Biosafety Level:2
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: 2000
Age: embryo, 10.5-days gestation
Comments:MEFs were derived from e10.5 embryos. 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. Both Mfn1 and Mfn1 are essential for mitochondrial fusion which is essential for embryonic development. These cell lines are useful in studying mitochondrial biology and the role of mitochondrial fusion in cell physiology.
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: bovine calf 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.

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 37.0°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 37.0°C.

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 30-2030
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-2994
Related cell line: ATCC CRL-2995
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