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Chronic Trafficking Assay

The chronic trafficking assay evaluates compound functional effects on ion channel protein trafficking in adult human iPSC cardiomyocytes (20 – 24 hour exposure) at physiological temperature. Compounds devoid of acute actions on cardiac ion channels may have significant inhibition of ion channel protein trafficking and ion channel function. Many drugs with cardiac toxicity inhibit ion channel protein trafficking. Compounds detected to have activity in this assay may possess cardiac risk, and can be further evaluated with individual ion channel studies if requested by the client. 
 
Standard Chronic Trafficking Protocol
  • Expose cell to drug for 24 hours
  • Record effects of drug exposure
Measurement Parameters
  • Resting Membrane Potential (mV)
  • Action Potential Amplitude (mV)
  • Action Potential Upstroke Velocity (V/s)
  • Action Potential Duration (ms)
  • Rate of Contraction (BPM - beats per minute)

chronic

A. Acute application of pentamidine at 1, 3, 10, and 30 μM (black) had no effect on the cardiac action potential; but prolongation, demonstrating the presence of IKr (hERG) current, was seen with 30 nM of the hERG blocker, E-4031 (red).

B. Action potential recording from a single ventricular-like adult human iPSC cardiomyocyte exposed to pentamidine for 24 hours, followed by perfusion with a drug-free external solution. Chronic pentamidine exposure increased the action potential duration (APD) and elicited early after-depolarizations.

 
 
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