Objective：To establish human induced pluripotent stem cell derived atrial and ventricular cardiomyocytes for cardiotoxicity assessment. Method：Temporarily manipulating retinoic acid signal to differentiate atrial and ventricular cardiomyocytes. Flow cytometry analysis and immunofluorescent staining were conducted to detect the cardiac-specific marker：α-actinin and ventricle-specific marker：MLC2v. qRT-PCR was done to detect ventricle-specific marker（MLC2v，MYH7）and atrial-specific marker（NR2F2，KCNA5）. Cell viability and calcium transient were assessed to evaluate cytotoxicity and cellular electrophysiological alterations caused by different drugs（terfenadine，sotalol and ibutilide）. Result：The results of flow cytometry，immunofluorescent staining and qRT-PCR demonstrated that ventricle-specific markers were highly expressed in RAi group while atrial-specific markers were expressed in RA group. By assessing calcium transient and cell viability，we proposed a drug assessment platform using hiPSC-ACM and hiPSC-VCM. After treated with terfenadine，cytotoxic effects occured at a concentration of 100 μmol/L both in hiPSC-ACM and hiPSC-VCM. Meanwhile，both sotalol and ibutilide exhibited cardiotoxicity potential at a concentration of 1 mmol/L. In the following calcium transient assessment，the amplitudes of calcium transient were significantly decreased in hiPSC-ACM after 1 μmol/L sotalol and 1 μmol/L ibutilide treatment，while in hiPSC-VCM，the amplitude showed no significant difference in such concentrations. The amplitudes of calcium transient were significantly decreased in both groups when the concentration of terfenadine reached to 1 mmol/L. Conclusion：By modulating retinoic acid signaling during hiPSC differentiation，we generated atrial and ventricular cardiomyocytes. Human induced pluripotent stem cell-derived atrial and ventricular cardiomyocytes were useful models for assessing cardiotoxicity of drugs by detecting calcium transient.