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SNLP 76/7-4 (ATCC® SCRC-1050™)

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

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.html.

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 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 embryonic stem cells [PubMed: 17576687; PubMed: 16236760] and induced pluripotent stem (iPS) cells [PubMed: 18079707]. Inclusion of puromycin in the medium is not necessary for normal cell growth.

ATCC tested that this cell line is resistant to:

  • G418 (neomycin):  350 µg/mL
  • Puromycin: 3 µg/mL

 
Storage Conditions liquid nitrogen vapor phase
Derivation
SNL76/7, ATCC SCRC-1049™ was clonally-derived from a STO cell line that expresses both G418 resistance and leukaemic inhibitory factor (LIF) at an abundant level [PubMed: 2205396].  SNLP 76/7-4, ATCC SCRC-1050™ is a puromycin resistant derivative of SNL76/7 [PubMed: 17576687].
Clinical Data
Male and female mixed
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.Remove and discard culture medium. 

  1. 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.
  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 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.
  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. An inoculum of 1 X 104 viable cells/cm2 is recommended
  5. Incubate cultures at 37°C. 
Subcultivation Ratio: 1:10

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
Storage Temperature: Liquid nitrogen vapor phase
Culture Conditions
Temperature: 37.0°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

Cadiñanos J, Bradley A. Generation of an inducible and optimized piggyBac transposon system. Nucleic Acids Res. 35(12): e87, 2007. PubMed: 17576687

Kist R, et al. Reduction of Pax9 gene dosage in an allelic series of mouse mutants causes hypodontia and oligodontia. Hum. Mol. Genet. 14(23): 3605-3617, 2005. PubMed: 16236760

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo JL. Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Basic Documentation
Product Sheet
Product Sheet
Certificate of Analysis
Certificate of Analysis
SDS
SDS
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

Cadiñanos J, Bradley A. Generation of an inducible and optimized piggyBac transposon system. Nucleic Acids Res. 35(12): e87, 2007. PubMed: 17576687

Kist R, et al. Reduction of Pax9 gene dosage in an allelic series of mouse mutants causes hypodontia and oligodontia. Hum. Mol. Genet. 14(23): 3605-3617, 2005. PubMed: 16236760

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo JL. Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.