Dicer1f/f (ATCC® CRL-3220)

Organism: Mus musculus, mouse  /  Cell Type: mesenchymal stem cell  /  Tissue: mesenchyme  / 

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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 Cell Micrograph of Dicer 1f/f ATCC CRL-3220
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.
  1. 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.
  2. 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.
  3. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
  4. 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

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • For-profit customers must obtain a research use license agreement from the Contributor prior to shipment.
  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
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Restrictions

This material is subject to the following restrictions in addition to those outlined in the ATCC Material Transfer Agreement:

  1. Prior to purchase, any for-profit Purchaser must obtain a research use license from Massachusetts Institute of Technology; and
  2. When a Commercial Use is contemplated, all for-profit and non-profit Purchasers must obtain a Commercial Use license from Massachusetts Institute of Technology.  Please contact Massachusetts Institute of Technology at the address listed below:
MIT Technology Licensing Office
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Technology Licensing Office, NE18-501
Cambridge, Massachusetts 02142
617-253-6966 tlo@mit.edu

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