CRL-1458 L6 大鼠成肌细胞 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 Permits/Forms: In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location. 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
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