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3T3 MEFs WT
3T3 MEFs WT
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編號(hào):B210943
品牌:Mingzhoubio

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產(chǎn)品名稱(chēng) 3T3 MEFs WT
商品貨號(hào) B210943
Organism Mus musculus, mouse
Tissue embryo
Cell Type fibroblast; spontanous immortalization (3T3)
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1

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 13.5 days gestation embryo
Applications
These cell lines are useful in studying the role of Caveolin-1 in a variety of signaling and membrane trafficking events.
Storage Conditions liquid nitrogen vapor phase
Images
Derivation
Mice homozygous null for the caveolin-1 gene, Cav-1 (-/-), and their wild-type littermates, Cav-1 (+/+), were generated by targeted disruption of the caveolin-1 gene. A construct was introduced into WW6 embryonic stem (ES) cells by electroporation to disrupt the Cav-1 locus. Mouse embryonic fibroblasts (MEFs) were obtained from day 13.5 littermate mouse embryos and immortalized using the 3T3 protocol. RefRazani B, et al. Caveolin-1 null mice are viable but show evidence of hyperproliferative and vascular abnormalities. J. Biol. Chem. 276: 38121-38138, 2001. PubMed: 11457855 
Comments

The 3T3 MEFs KO cell line (ATCC CRL-2753) is homozygous for a disruption of the caveolin-1 gene Cav-1 (-/-) while the 3T3 MEFs WT cell line (ATCC CRL-2752) is Cav-1 (+/+). Analysis of cultured fibroblasts from Cav-1 null embryos reveals a loss of caveolin-2 protein expression; defects in the endocytosis of a known caveolar ligand, (fluorescein isothiocyanate-albumin); and a hyperproliferative phenotype. These phenotypic changes are reversed by recombinant expression of the caveolin-1 cDNA. RefRazani B, et al. Caveolin-1 null mice are viable but show evidence of hyperproliferative and vascular abnormalities. J. Biol. Chem. 276: 38121-38138, 2001. PubMed: 11457855

A culture deposited with the ATCC in September of 2002 was found to be contaminated with mycoplasma. Progeny were cured by a 21-day treatment with BM Cyclin. The cells were assayed for mycoplasma, by the Hoechst stain, PCR and the standard culture test, after a six-week period following treatment. All tests were negative.

Analysis of cultured fibroblasts from Cav-1 null embryos reveals a loss of caveolin-2 protein expression; defects in the endocytosis of a known caveolar ligand, (fluorescein isothiocyanate-albumin); and a hyperproliferative phenotype.

 

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 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Note:Subculture at 80% confluency.

  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum that 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 pipetting gently.
  5. Add appropriate aliquots of cell suspension to new culture vessels.
  6. Incubate culture vessels at 37°C.

Split Ratio: 1:5 to 1:10
Medium Renewal: Every 2 to 3 days.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture of Animal Cells: a Manual of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Name of Depositor MP Lisanti
References

Razani B, et al. Caveolin-1 null mice are viable but show evidence of hyperproliferative and vascular abnormalities. J. Biol. Chem. 276: 38121-38138, 2001. PubMed: 11457855

Sotgia F, et al. Intracellular retention of glycosylphosphatidyl inositol-linked proteins in caveolin-deficient cells. Mol. Cell. Biol. 22: 3905-3926, 2002. PubMed: 11997523

梅經(jīng)理 17280875617 1438578920
胡經(jīng)理 13345964880 2438244627
周經(jīng)理 17757487661 1296385441
于經(jīng)理 18067160830 2088210172
沈經(jīng)理 19548299266 2662369050
李經(jīng)理 13626845108 972239479
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