This cell line was derived as a subclone from the cloned cell strain of BD1X rat embryonic heart tissue by Kimes B and Brandt B; it exhibits many characteristics of skeletal muscle. The myoblasts in this cell line can fuse to form multinucleated myotubes and respond to stimulation by acetylcholine. When the serum concentration in the culture medium is reduced to 1%, fusion occurs rapidly.
Passage Ratio: 1:3-1:4, with medium renewal every 2-3 days Doubling Time: 48-72 h Culture Conditions: Atmosphere: 95% air + 5% CO₂, Temperature: 37°C
Freezing Conditions: 60% basal medium + 30% FBS + 10% DMSO, stored in liquid nitrogen
Recommended HyCyte® One-Step Cryopreservation Medium (ready-to-use, serum-free, no programmed cooling required), Product No.: GUCP-R201 Quality Control: Negative for bacteria, fungi, and mycoplasma detection
Abnormal Conditions in H9C2 Cell Culture and Preventive Measures:
(1) Excessive Granules on Cell Surface This may indicate issues with the culture environment. Switching to DMEM/F-12 medium can help, and the cell surface will become smooth again after three passages.
(2) Formation of Vacuoles This could be due to uneven plating, where localized overcrowding leads to poor cell condition and increased vacuolation. Alternatively, it may be caused by low-quality serum. Replace it with high-quality serum and change the medium promptly.
(3) Slow Cell Growth This might result from insufficient cell seeding or low cell density. Continue culturing, then digest and replate the cells at a higher density.
(4) Deteriorating Cell Condition and Detachment This could be due to serum issues. It is recommended to replace the serum and ensure it is thawed properly (programmed thawing, not water bath). Overcrowding may also be a cause. Timely passaging is essential—do not wait until the cells are fully confluent. Passage should be performed when localized overgrowth or overlapping occurs.