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微信小章| 产品名称： | 3T3 MEFs WT |
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| 商品货号： | TS165974 |
| 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. Ref   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: 11457855xa0 |
| 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 (TS165974) 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. Ref 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.
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| Complete Growth Medium： | The base medium for this cell line is ATCC-formulated Dulbeccos Modified Eagles 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%.
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| 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.
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 |