H9c2(2-1)

H9c2(2-1) cells, derived from the ventricular myoblasts of embryonic BD1X rat hearts, are a subclone of the original H9 cell line established in the early 1990s. These cells are immortalized myoblasts that are commonly used in vitro to study cardiac metabolism, physiology, and pathophysiology, including myocardial ischemia, hypertrophy, and apoptosis mechanisms.

Phenotypically, H9c2 cells exhibit characteristics of skeletal muscle but retain the ability to adopt a cardiac muscle phenotype under specific experimental conditions, such as differentiation induced by retinoic acid or other agents. This flexibility makes them a valuable model for investigating cardiac muscle behavior in response to various physiological and pharmacological stimuli. Genetically, H9c2 cells are diploid, facilitating their use in genetic studies, where maintaining a stable karyotype is crucial.

Research employing H9c2(2-1) cells has contributed significantly to understanding cellular responses to oxidative stress, mitochondrial dysfunction, and the protective roles of various pharmacological agents against cardiotoxicity. This cell line remains a cornerstone in cardiomyocyte-related research, offering a reproducible, controlled model to elucidate the complex biological and molecular mechanisms underlying cardiac function and diseases.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.