產(chǎn)品名稱 |
Dicer1f/f |
商品貨號(hào) |
B217772 |
Organism |
Mus musculus, mouse |
Tissue |
mesenchyme |
Cell Type |
mesenchymal stem cell |
Product Format |
frozen 1.0 mL |
Morphology |
mesenchymal-like |
Culture Properties |
adherent |
Biosafety Level |
2 [Cells contain SV40 viral DNA sequences]
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 month old |
Gender |
female |
Strain |
Dicer1tm1Bdh/J |
Applications |
Dicer 1f/f cells (ATCC® CRL-3220™) are the wild-type control for the Dicer1 -/- mesenchymal stem cells (ATCC® CRL-3221™). Dicer 1f/f cells can be used in conjunction with the matching Dicer1-/- cells to study the role of Dicer and miRNA in cell biology. |
Storage Conditions |
liquid nitrogen vapor phase |
Images |
 |
Derivation |
Primary mesenchymal stem cell (MSC) cultures were prepared from the tibia, femur, and pelvic bones of a one-year old Dicer f/f mouse. The primary cells were then infected with retrovirus encoding SV40 large T-antigen, grown out, and subcloned into individual monoclonal populations. |
Comments |
MicroRNAs (miRNAs) are a new class of small noncoding RNAs that post-transcriptionally regulate the expression of target mRNA transcripts. Dicer is an RNase III family endoribonuclease that has the biofunction of processing miRNAs. Loss of Dicer1 leads to a dramatic decrease in levels of cellular microRNA, allowing for investigations into the roles of microRNAs in various cellular functions.
Dicer 1f/f (ATCC® CRL-3220™) is the wild-type control for Dicer 1-/- (ATCC® CRL-3221™), Dicer deficient mesenchymal stem cells. Dicer is an RNase III family endoribonuclease tumor suppressor that processes miRNAs. It has been suggested that complete Dicer 1 loss and the subsequent misregulation of gene expression can inhibit tumor growth rates through reduced proliferation and increased cell death. This suggests that the targeted inhibition of miRNA pathway elements, particularly Dicer I, may be a potential therapy for the treatment of cancer. |
Complete Growth Medium |
Alpha minimum essential medium with ribonucleosides and deoxyribonucleosides, 90%; fetal bovine serum, 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.
- Remove and discard culture medium. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) (ATCC® 30-2200) or 0.25% Trypsin – 0.53mM EDTA (ATCC® 30-2101) solution to remove all traces of serum which 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: 1:3 to 1:6 is recommended.
Medium renewal 2 to 3 times a week |
Cryopreservation |
Freeze Medium: Fetal Bovine Serum (FBS), 92%; DMSO, 8%
Storage Temperature: liquid nitrogen vapor phase |
Culture Conditions |
Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5% |
Volume |
1.0 mL |
Name of Depositor |
P. Sharp, Massachusettes Institute of Technology |
Year of Origin |
2009 |
References |
Ravi A, et al. Proliferation and tumorigenesis of a murine sarcoma cell line in the absence of DICER1. Cancer Cell 21(6): 848-855, 2012. PubMed: 22698408
Gurtan AM, et al. In vivo structure-function analysis of human Dicer reveals directional processing of precursor miRNAs. RNA 18: 1116-1122, 2012. PubMed: 22546613
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