SNL76/7 (ATCC® SCRC-1049)

Organism: Mus musculus, mouse  /  Cell Type: Fibroblast  /  Tissue: Embryo  / 

Permits and Restrictions

View Permits View Restrictions

Organism Mus musculus, mouse
Tissue
Embryo
Cell Type Fibroblast
Product Format frozen
Morphology Fibroblast
Culture Properties Adherent
Biosafety Level 1 Appropriate safety procedures should always be used with this material. Laboratory safety is discussed in the following publication: Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS Publication No. (CDC) 93-8395. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at

http://www.cdc.gov/biosafety/publications/bmbl5/index.htm.

Age embryo
Gender Male and female mixed
Strain SIM (Sandos Inbred Mice)
Applications
This cell line can be used as a feeder layer to support the growth of mouse embryonic stem cells and induced pluripotent stem (iPS) cells.
Storage Conditions liquid nitrogen vapor phase
Derivation
SNL76/7 was clonally-derived from a STO cell line that expresses both G418 resistance and leukaemic inhibitory factor (LIF) at an abundant level.
Clinical Data
Male and female mixed
Comments

The STO cell line was transfected with a G418-resistance cassette, RV4.0 [PubMed: 2822260] and a LIF expression construct [PubMed: 3143916]. Inclusion of G418 in the medium is not necessary for normal cell growth. Note: ATCC tested that this cell line is resistant to: G 418 (neomycin): 350 µg/mL.

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%.
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.

  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum, which contains trypsin inhibitor.
  3. 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 10 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. 
  5. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 1 X 104 viable cells/cm2 is recommended.
  6. Incubate cultures at 37°C.
Subcultivation Ratio: 1:10

Interval: every 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

Complete growth medium supplemented with an additional 35% fetal bovine serum and 10% (v/v) DMSO.

Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

Culture Conditions
Temperature: 37°C
Atmosphere: Air, 95%; carbon dioxide (CO2), 5%
Name of Depositor A Bradley
Year of Origin 1988
References

McMahon AP, Bradley A. The Wnt-1 (int-1) proto-oncogene is required for development of a large region of the mouse brain. Cell. 62(6): 1073-1085, 1990. PubMed: 2205396

Takahashi K, et al. Induction of pluripotent stem cells from fibroblast cultures. Nat. Protoc. 2(12): 3081-3089, 2007. PubMed: 18079707

Thomas KR, Capecchi MR. Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell 51(3): 503-512, 1987. PubMed: 2822260

Williams RL, et al. Myeloid leukaemia inhibitory factor maintains the developmental potential of embryonic stem cells. Nature 336: 684-687, 1988. PubMed: 3143916

Okita K, et al, Generation of germline-competent induced pluripotent stem cells. Nature 448(7151): 313-317, 2007. PubMed: 17554338

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • 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.
Basic Documentation
Restrictions

Prior to purchase, for-profit commercial institutions must obtain a license agreement. For instructions on how to proceed, please contact Baylor Licensing Group, blg@bcm.tmc.edu

References

McMahon AP, Bradley A. The Wnt-1 (int-1) proto-oncogene is required for development of a large region of the mouse brain. Cell. 62(6): 1073-1085, 1990. PubMed: 2205396

Takahashi K, et al. Induction of pluripotent stem cells from fibroblast cultures. Nat. Protoc. 2(12): 3081-3089, 2007. PubMed: 18079707

Thomas KR, Capecchi MR. Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell 51(3): 503-512, 1987. PubMed: 2822260

Williams RL, et al. Myeloid leukaemia inhibitory factor maintains the developmental potential of embryonic stem cells. Nature 336: 684-687, 1988. PubMed: 3143916

Okita K, et al, Generation of germline-competent induced pluripotent stem cells. Nature 448(7151): 313-317, 2007. PubMed: 17554338