3T3 MEFs WT細胞
ATCC Number： CRL-2752?
數量： 大量
細胞類型： 其他細胞類型
運輸方式： 凍存運輸
細胞形態： 成纖維樣
是否是腫瘤細胞： 0
物種來源： 小鼠
生長狀態： 貼壁生長
器官來源： 胚胎
3T3 MEFs WT細胞年限： 13.5 days gestation embryo
Designations: 3T3 MEFs WT
Depositors: MP Lisanti
Biosafety Level: 1
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Mus musculus
Morphology: fibroblast
Source: Organ: embryo
Cell Type: fibroblast fibroblast; spontanous immortalization (3T3)
Permits/Forms: 3T3 MEFs WT細胞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.
Age: 13.5 days gestation embryo
Comments: 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 [PubMed: 11457855]. 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 [PubMed: 11457855].These cell lines are useful in studying the role of Caveolin-1 in a variety of signaling and membrane trafficking events. 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.
Propagation: 3T3 MEFs WT細胞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%.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Subculturing: Protocol: Subculture at 80% confluency.
Remove and discard culture medium.
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.
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: 3T3 MEFs WT細胞A subcultivation ratio of 1:5 to 1:10 is recommended
Medium Renewal: Every 2 to 3 days
Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002
recommended serum:ATCC 30-2020
References: 60275: 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
60276: Sotgia F, et al. Intracellular retention of glycosylphosphatidyl inositol-linked proteins in caveolin-deficient cells. Mol. Cell. Biol. 22: 3905-3926, 2002. PubMed: 11997523
