AC16 Cardiomyocyte Cell Line

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.

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).

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.