AC16 human cardiomyocyte cell lines were purchased from EMD Millipore (cod. SCC109). Following the manufacturer's instructions, the cell line was tested and authenticated for mycoplasma contamination, resulting in negative data. Cells were cultured in Dulbecco's Modified Eagle's Medium /Nutrient Mixture F-12 (cod. D8437, Sigma) containing 12.5% fetal bovine serum (FBS) (cod. ECS0180L, Euroclone), 1% antibiotics penicillin–streptomycin (cod. ECB3001D, Euroclone), and 1% of L-glutamine (cod. ECB3000D, Euroclone). The cell line was maintained in the incubator at 37 °C and 5% CO2. The cells were grown between 4 and 6 passages, and experiments were performed in triplicate. AC16 were exposed to 33 mmol/L D glucose (cod. G8644, EMD Millipore) for 7 days [19 (link)] and treated with tirzepatide (LY3298176, selleckchem) at a concentration of 100 nM. The medium was changed every 48 h. Normal glucose, NG, was considered the cells exposed to normal glucose concentration (5.5 mmol/L) and cultured for 7 days. A dose–response curve, using cell viability and toxicity assay, was performed to evaluate the right concentration of tirzepatide to carry out experiments (Additional file 1 : figure).
Tirzepatide
Manufactured by Selleck Chemicals
Tirzepatide is a synthetic peptide molecule developed for use in laboratory research. It functions as a dual agonist, binding and activating both the glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) receptors.
Automatically generated - may contain errors
Lab products found in correlation
Ecb3001d, by Euroclone (1 mentions)
G8644, by Merck Group (1 mentions)
Ecs0180l, by Euroclone (1 mentions)
Ecb3000d, by Euroclone (1 mentions)
Forskolin, by Merck Group (1 mentions)
Exendin 4, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Glomax discover microplate reader, by Promega (1 mentions)
Ultrasensitive rat insulin elisa, by Crystal Chem (1 mentions)
2 protocols using tirzepatide
1
High Glucose-Induced Cardiomyocyte Dysfunction: Tirzepatide Evaluation
2
Glucose-stimulated Insulin Secretion Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The method has been previously described [12 (link)]. In brief, pancreatic islets were first starved in Krebs Ringer HEPES (KRH) buffer supplemented with 2 mM glucose ± Kom56. Subsequently, pancreatic islets were incubated in fresh KRH buffer ± Kom56 containing 2 mM glucose (low glucose) to measure the basal insulin secretion, and afterwards, the same pancreatic islets were incubated in fresh KRH buffer ± Kom56 containing 20 mM glucose (high glucose) to measure the glucose-stimulated insulin secretion (GSIS). For the insulin measurement after treatment with different glucose concentrations (2 mM, 5 mM, 10 mM, and 15 mM), the pancreatic islets were first starved in KRH buffer supplemented with 2 mM glucose ± Kom56. To determine the insulin secretion, the same pancreatic islets were subsequently incubated with fresh KRH buffer ± Kom56 and containing the respective glucose concentration. Kom56 and exendin-4 (Sigma–Aldrich) were dissolved in water, forskolin (Sigma–Aldrich) and tirzepatide (Selleckchem) were dissolved in DMSO (Sigma–Aldrich). Insulin concentrations were measured using an ultra-sensitive rat insulin ELISA (Crystal Chem) in combination with a GloMax Discover Microplate Reader (Promega).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!