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Dicer1f/f

CRL-3220

Product category
Animal cells
Organism
Mus musculus, mouse
Classification
Eukaryota, Animalia, Metazoa, Chordata, Vertebrata, Tetrapod
Cell type
mesenchymal stem cell
Morphology
mesenchymal-like
Tissue
Embryo; Mesenchyme
Applications
3D cell culture
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
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Price: $844.00 EA
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Documentation

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

Cells contain SV40 sequences

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Required Products

These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.

Detailed product information

General

Specific 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.

Characteristics

Volume
1.0 mL
Growth properties
Adherent
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.
Age
13 months
Gender
Female
Strain
Dicer1tm1Bdh/J
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.

Handling information

Unpacking and storage instructions
  1. Check all containers for leakage or breakage.
  2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete medium
Alpha minimum essential medium with ribonucleosides and deoxyribonucleosides, 90%; fetal bovine serum, 10%
Temperature
37°C
Atmosphere
95% Air, 5% CO2
Handling procedure

To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.  Storage at -70°C will result in loss of viability.  

  1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water.  Thawing should be rapid (approximately 2 minutes).
  2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
  3. Transfer the vial contents to a centrifuge tube containing  9.0 mL complete culture medium. and spin at approximately 125 x g for 5 to 7 minutes.
  4. Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). It is important to avoid excessive alkalinity of the medium during recovery of the cells.  It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6). pH (7.0 to 7.6).
  5. Incubate the culture at 37°C in a suitable incubator.  A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet.

         

Subculturing procedure
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
Reagents for cryopreservation
Fetal Bovine Serum supplemented with 8% (v/v) DSMO (ATCC 4-X)

History

Depositors
P. Sharp, Massachusettes Institute of Technology
Year of origin
2009

Legal disclaimers

Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Warranty

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Permits & Restrictions

For-profit Research Use License from Massachusetts Institute of Technology

For-profit Organizations: For every order of this item, you must work directly with the contributor, Massachusetts Institute of Technology to (i) negotiate a research-use license, and/or (ii) have the contributor provide authorization to ATCC to ship this item under your existing license. We cannot ship this item until we receive communication directly from Massachusetts Institute of Technology that we are authorized to ship each order.

We are providing the following contact information, but this information may change without notice:
MIT Technology Licensing Office
One Cambridge Center, Kendall Square
Technology Licensing Office, NE18-501
Cambridge, Massachusetts 02142
Phone: 617-253-6966
Email: [email protected]

Once ATCC has received authorization from Massachusetts Institute of Technology, your order will be reviewed, and this item will be released for shipment if all requirements are met. If you need assistance with your order, please contact our Customer Care team or your applicable distributor.

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

Frequently Asked Questions

References

Curated Citations

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

For product-related inquiries and issues, contact Technical Service:

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