H9c2(2-1) (ATCC® CRL-1446)

Organism: Rattus norvegicus, rat  /  Cell Type: Myoblast  /  Tissue: heart/myocardium  / 

Organism Rattus norvegicus, rat
Tissue heart/myocardium
Cell Type Myoblast
Product Format frozen
Morphology myoblast
Culture Properties adherent
Biosafety Level 1
Age embryo
Strain BD1X
Applications
This cell line is a suitable transfection host.
Storage Conditions liquid nitrogen vapor phase
Derivation
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.
Receptor Expression
acetylcholine, expressed
Genes Expressed
myokinase; creatine phosphokinase; myosin
Cellular Products
myokinase; creatine phosphokinase; myosin
Comments
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.
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. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.

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.
  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 which 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 1:2 to 1:4 is recommended
Medium Renewal: Every 2 to 3 days
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 W Carlisle
References

Kimes BW, Brandt BL. Properties of a clonal muscle cell line from rat heart. Exp. Cell Res. 98: 367-381, 1976. PubMed: 943302

Levy AP, et al. Post-transcriptional regulation of vascular endothelial growth factor by hypoxia. J. Biol. Chem. 271: 2746-2753, 1996. PubMed: 8576250

Basic Documentation
FAQ's
  1. CRL-1446 culture medium and cell density


    Date Updated: 2/20/2014

References

Kimes BW, Brandt BL. Properties of a clonal muscle cell line from rat heart. Exp. Cell Res. 98: 367-381, 1976. PubMed: 943302

Levy AP, et al. Post-transcriptional regulation of vascular endothelial growth factor by hypoxia. J. Biol. Chem. 271: 2746-2753, 1996. PubMed: 8576250