| Field | Specification |
|---|---|
| Species |
The AC16 cell line, derived from human ventricular cells fused with SV40-transformed, showcases characteristics typical of cardiomyocytes, including the expression of transcription factors such as GATA4, MYCD, NFATc4, and contractile proteins like alpha- and beta-myosin heavy chain. AC16 cells also express gap junction proteins connexin-43 and connexin-40, with functional gap junctions confirmed by dye-coupling studies, underscoring their utility in cardiomyocyte research. When the SV40 oncogene is silenced, AC16 transitions towards a more differentiated state, marked by the expression of BMP2, indicative of cardiac differentiation and developmental regulation.
In general, scientists employ various techniques, including stem cell differentiation, animal models, molecular analysis, and biomarker discovery, to advance knowledge and potential therapies for heart-related conditions. The involvement of mitogen and senescence pathways, along with thymidine kinase induction, further elucidates the complex nature of human cardiomyocytes and their response to pathological conditions.
The AC16 human cardiomyocyte cell line's ability to mimic the behavior of mature cardiomyocytes makes it a valuable model for cardiac research. It closely resembles the genetic makeup of primary cardiomyocytes, allowing for studies on cardiac development, pathology, and the implications of histone loss in vitro, however, the cardiomyocyte behavior and genetic complexity might not fully match that of primary or stem cell-derived cardiomyocytes. In the context of toxicology and cardiovascular disease research, AC16 cells serve as a vital tool for understanding cardiomyocyte development, inflammation, injury, regeneration, and toxicological effects.
The unique properties of the AC16 human cardiomyocyte cell line, including its response to developmental cues and the ability to simulate the physiological conditions of human cardiomyocytes, make it an indispensable asset in the quest to unravel the mysteries of heart diseases and devise novel therapeutic interventions.
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Viruses: Transformed by the SV40 large T-antigen
- cultureMedium:
- Culture medium:DMEM:Ham's F12 (1:1), w: 3.1 g/L Glucose, w: 2.5 mM L-Glutamine, w: 15 mM HEPES, w: 0.5 mM Sodium pyruvate, w: 1.2 g/L NaHCO3 (Cytion article number 820400a). Supplement the culture medium with 12.5% FBS and add 0.9 mM L-Glutamine to achieve a final concentration of 2.5 mM L-Glutamine.
- Differentiation medium: DMEM:Ham's F12 (1:1), w: 3.1 g/L Glucose, w: 2.5 mM L-Glutamine, w: 15 mM HEPES, w: 0.5 mM Sodium pyruvate, w: 1.2 g/L NaHCO3 (Cytion article number 820400a). To prepare the complete differentiation medium, add 1x ITS+ (Gibco, catalog number 41400045) and 2% Horse Serum (Gibco, catalog number 16050130).
- dissociationReagent: Accutase
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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