Normal h9c2 cardiomyocyte cell line

As we previously described [13 (link)], neonatal rat ventricular myocytes (NRVMs) were obtained from isolated heart tissues of young Wistar rats. Briefly, the dissected hearts minced in HEPES-buffered nic acid saline solution. Non-myocyte contaminants were removed by two rounds of pre-plating for 1.5 h on 100-mm plastic cell culture dishes under a culture condition of 37 °C and 5% CO₂. Then, the cardiomyocytes were seeded into different culture dishes with medium containing serum. After incubation for 24 hours, medium without serum was used to replace the serum-containing medium.
The normal H9c2 cardiomyocyte cell line was purchased from American Type Culture Collection (ATCC). Cells were cultured with 1640 medium containing 10% fetal bovine serum in a humidified atmosphere of 5% CO₂ at 37 °C. Actinomycin D was purchased from Sigma-Aldrich (St. Louis, MO, USA) and used at the concentration of 5 μg/ml. MCC950 sodium (CP-456773, CRID3 sodium salt), a NLRP3 inhibitor, was used at the concentration of 7.0 nM in cell culture and 12 mg/kg in animal treatment (Selleck, Shanghai, China). For osmotic stress, hyperosmotic medium was made by adding various concentrations of D-sorbitol (Sigma-Aldrich) to regular 1640 medium for different durations. Standard 1640 culture condition was used as isosmotic media.