L6 (ATCC® CRL-1458)

Organism: Rattus norvegicus, rat  /  Cell Type: myoblast  /  Tissue: skeletal muscle  / 

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Organism Rattus norvegicus, rat
Tissue skeletal muscle
Cell Type myoblast
Product Format frozen
Morphology myoblast
Culture Properties adherent
Biosafety Level 1
Applications
This cell line is a suitable transfection host.
Storage Conditions liquid nitrogen vapor phase
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Derivation
The L6 myogenic line was isolated originally by Yaffe from primary cultures of rat thigh muscle maintained for the first two passages in the presence of methyl cholanthrene.
Genes Expressed
myosin
Comments
L6 cells fuse in culture to form multinucleated myotubes and striated fibers. The extent of cell fusion declines with passage and the cells should be frozen at low passage and periodically recloned with selection for fusion competent cells.
Tested and found negative for ectromelia virus (mousepox).
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 are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes. Subculture before the cells become confluent to retard the loss of differentiating ability that is observed as the cells are passaged.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that 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 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.
  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.
  6. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:20 to 1:40 is recommended
Medium Renewal: 2 to 3 times per week
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Growth Conditions: The myoblastic component of this line will be depleted rapidly if the cells are allowed to become confluent.
Name of Depositor D Schubert
Passage History
The L6 myogenic line was isolated originally by Yaffe from primary cultures of rat thigh muscle maintained for the first two passages in the presence of methyl cholanthrene.
L6 cells fuse in culture to form multinucleated myotubes and striated fibers. The extent of cell fusion declines with passage and the cells should be frozen at low passage and periodically recloned with selection for fusion competent cells.
References

Mandel JL, Pearson ML. Insulin stimulates myogenesis in a rat myoblast line. Nature 251: 618-620, 1974. PubMed: 4421831

Richler C, Yaffe D. The in vitro cultivation and differentiation capacities of myogenic cell lines. Dev. Biol. 23: 1-22, 1970. PubMed: 5481965

Yaffe D. Retention of differentiation potentialities during prolonged cultivation of myogenic cells. Proc. Natl. Acad. Sci. USA 61: 477-483, 1968. PubMed: 5245982

Osawa H, et al. Identification and characterization of basal and cyclic AMP response elements in the promoter of the rat hexokinase II gene. J. Biol. Chem. 271: 17296-17303, 1996. PubMed: 8663388

Osawa H, et al. Analysis of the signaling pathway involved in the regulation of hexokinase II gene transcription by insulin. J. Biol. Chem. 271: 16690-16694, 1996. PubMed: 8663315

Cross References

Nucleotide (GenBank) : M87067 R.norvegicus activin type IIB receptor mRNA.

Nucleotide (GenBank) : X57986 Rat mRNA for C alpha subunit of the cAMP-dependent protein kinase.

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
Other Documentation
FAQ's
  1. CRL-1458 culture conditions and cell density
    CRL-1458 cells are confluence sensitive.  These cells should be seeded at an initial density of 2 X 104 viable cells...
    Date Updated: 2/20/2014
References

Mandel JL, Pearson ML. Insulin stimulates myogenesis in a rat myoblast line. Nature 251: 618-620, 1974. PubMed: 4421831

Richler C, Yaffe D. The in vitro cultivation and differentiation capacities of myogenic cell lines. Dev. Biol. 23: 1-22, 1970. PubMed: 5481965

Yaffe D. Retention of differentiation potentialities during prolonged cultivation of myogenic cells. Proc. Natl. Acad. Sci. USA 61: 477-483, 1968. PubMed: 5245982

Osawa H, et al. Identification and characterization of basal and cyclic AMP response elements in the promoter of the rat hexokinase II gene. J. Biol. Chem. 271: 17296-17303, 1996. PubMed: 8663388

Osawa H, et al. Analysis of the signaling pathway involved in the regulation of hexokinase II gene transcription by insulin. J. Biol. Chem. 271: 16690-16694, 1996. PubMed: 8663315