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Use of human iPSC-derived cardiomyocytes to monitor compound effects on pathways
Cancer researchers in Maryland were able to use human induced pluripotent stem cell derived-cardiomyocytes (hiPSC-CMs) to investigate compound effects on cardiac myocyte signaling pathways. Their goal was to further demonstrate that hiPSC-CMs were a suitable in vitro model for studies at the molecular level, and to lay out workflows involving a variety of interdependent endpoints from maintenance of cell cultures to characterizing biological consequences of signaling modulation. They have published their comprehensive guidelines seeking to encourage more utilization of hiPSC-CMs for mechanistic investigations.
To align with their goals of advancing studies in cardiac myocyte function the researchers chose to put forth a robust method that could help pinpoint which signaling pathway to focus on. In their set of basic protocols is a thorough description of a multiparameter biochemical and imaging assay optimized to evaluate overall cell viability. Such an assay would prove useful in determining how cells are affected by certain conditions. The assay's parameters included analysis of nuclear morphology, mitochondrial membrane potential, and markers towards apoptotic events, which was assessed using materials such as Hoechst 33258, Cell Meter™ Mitochondrion Membrane Potential JC-10 Assay Kit with CCCP, and a human cardiac troponin T assay kit, respectively.
In addition to the materials list, the researchers also had explanations for how each item worked, as well as justification of why they designed the assay to be as it was. They emphasized that their overall approach "saves time and expense", which is interesting to note because the Cell Meter™ Mitochondrion Membrane Potential JC-10 Assay Kit was developed with the exact same intent. The Cell Meter™ JC-10 Kit has all reagents and instructions prepared for swift and convenient monitoring of mitochondrial membrane potential. The fluorescent probe JC-10 is an ideal dye for in vitro studies, including ones involving iPSC-derived cells, due to its high water solubility.
To align with their goals of advancing studies in cardiac myocyte function the researchers chose to put forth a robust method that could help pinpoint which signaling pathway to focus on. In their set of basic protocols is a thorough description of a multiparameter biochemical and imaging assay optimized to evaluate overall cell viability. Such an assay would prove useful in determining how cells are affected by certain conditions. The assay's parameters included analysis of nuclear morphology, mitochondrial membrane potential, and markers towards apoptotic events, which was assessed using materials such as Hoechst 33258, Cell Meter™ Mitochondrion Membrane Potential JC-10 Assay Kit with CCCP, and a human cardiac troponin T assay kit, respectively.
In addition to the materials list, the researchers also had explanations for how each item worked, as well as justification of why they designed the assay to be as it was. They emphasized that their overall approach "saves time and expense", which is interesting to note because the Cell Meter™ Mitochondrion Membrane Potential JC-10 Assay Kit was developed with the exact same intent. The Cell Meter™ JC-10 Kit has all reagents and instructions prepared for swift and convenient monitoring of mitochondrial membrane potential. The fluorescent probe JC-10 is an ideal dye for in vitro studies, including ones involving iPSC-derived cells, due to its high water solubility.
References
- Guo, L. , Eldridge, S. , Furniss, M. , Mussio, J. , and Davis, M. 2015. Use of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) to monitor compound effects on cardiac myocyte signaling pathways. Curr. Protoc. Chem. Biol. 7:141-185. doi: 10.1002/9780470559277.ch150035
- Cell Meter™ JC-10 Mitochondrion Membrane Potential Assay Kit. AAT Bioquest, n.d. Web. 24 June 2016
Original created on January 3, 2020, last updated on January 3, 2020
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