產品名稱 |
L6 |
商品貨號 |
B164949 |
Organism |
Rattus norvegicus, rat |
Tissue |
skeletal muscle |
Cell Type |
myoblast |
Product Format |
frozen |
Morphology |
myoblast |
Culture Properties |
adherent |
Biosafety Level |
1
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. |
Applications |
This cell line is a suitable transfection host. |
Storage Conditions |
liquid nitrogen vapor phase |
Images |
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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 |
Cellular Products |
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%.
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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. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.
- Remove and discard culture medium.
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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.
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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.
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Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
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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
Note: The myoblastic component of this line will be depleted rapidly if the cells are allowed to become confluent. |
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 |
Name of Depositor |
D Schubert |
Deposited As |
Rattus sp. |
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
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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.
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