CRL-1458 L6 大鼠成肌細胞
ATCC® Number: CRL-1458?
Designations: L6
Depositors: D Schubert
Biosafety Level: 1
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Rattus norvegicus (rat)
Morphology: myoblast
Source: Tissue: skeletal muscle
Cell Type: myoblast myoblast;
Cellular Products: myosin
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Applications: transfection host (Nucleofection technology from Lonza
Roche FuGENE® Transfection Reagents)
Comments: 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. [22581]
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).
Propagation: ATCC 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%.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Growth Conditions: The myoblastic component of this line will be depleted rapidly if the cells are allowed to become confluent.
Subculturing: Protocol: Subculture before the cells become confluent to retard the loss of differentiating ability that is observed as the cells are passaged.
Remove and discard culture medium.
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.
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.
Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
Add appropriate aliquots of the cell suspension to new culture vessels.
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
Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002
recommended serum:ATCC 30-2020
References: 1064: Mandel JL, Pearson ML. Insulin stimulates myogenesis in a rat myoblast line. Nature 251: 618-620, 1974. PubMed: 4421831
22255: Richler C, Yaffe D. The in vitro cultivation and differentiation capacities of myogenic cell lines. Dev. Biol. 23: 1-22, 1970. PubMed: 5481965
22581: Yaffe D. Retention of differentiation potentialities during prolonged cultivation of myogenic cells. Proc. Natl. Acad. Sci. USA 61: 477-483, 1968. PubMed: 5245982
33164: 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
33165: 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
