ATCC® Number: CRL-1446™ Designations: H9c2（2-1） Depositors: W Carlisle Biosafety Level: 1 Shipped: frozen Medium & Serum: See Propagation Growth Properties: adherent Organism: Rattus norvegicus （rat） Morphology: myoblast
Source: Strain: BD1X Organ: heart Tissue: myocardium Cellular Products: myokinase; creatine phosphokinase; 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 ultimay responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Applications: transfection host （Roche FuGENE® Transfection Reagents） Receptors: acetylcholine, expressed Age: embryo Comments: H9c2（2-1） is a subclone of the original clonal cell line derived from embryonic BD1X rat heart tissue by B. Kimes and B. Brandt and exhibits many of the properties of skeletal muscle. Myoblastic cells in this line will fuse to form multinucleated myotubes and respond to acetylcholine stimulation. Fusion occurs faster if the serum concentration in the medium is reduced to one percent. 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 ?H9c2（2-1） 大鼠心肌細胞 Subculturing: Protocol: The myoblastic population will become depleted rapidly if the cultures are allowed to become confluent. To prevent loss of myoblastic cells, cultures should be subcultured before they become confluent, and the line should be recloned periodically with selection for myoblastic cells.
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 which 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.
Subc*tion Ratio: A subc*tion ratio of 1:2 to 1:4 is recommended Medium Renewal: Every 2 to 3 days 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 ?H9c2（2-1） 大鼠心肌細胞 References: 1062: Kimes BW, Brandt BL. Properties of a clonal muscle cell line from rat heart. Exp. Cell Res. 98: 367-381, 1976. PubMed: 943302 32970: Levy AP, et al. Post-transcriptional regulation of vascular endothelial growth factor by hypoxia. J. Biol. Chem. 271: 2746-2753, 1996. PubMed: 8576250
