Exploring stem cell-derived cardiomyocytes with automated patch clamp techniques


Biophysics 2012


Rikke P Schrøder, Mette T Christensen, Blake Anson, Morten Sunesen



There is increasing interest for cardiomyocytes as models for studying cardiac cellular physiology and preclinical drug safety testing. Stem cell-derived cardiomyocytes have the potential for such a model and have the possibility for modeling human diseases. The present investigation is the first to describe current properties from stem cell-derived cardiomyocytes using multi-hole recordings with planar automated patch clamp technology. In our study pluripotent stem cell-derived cardiomyocytes were biophysically and pharmacologically characterized. The cells are differentiated in large numbers and cryo-preserved, which make them suitable for automated patch clamping and facilitate their use in drug screening. We tested the cells in two different recording modes; single-hole and multi-hole, respectively. For multi-hole recordings up to ten cells are patched at the same time and the total current is measured per site. This recording mode can be useful for small currents (e.g. endogenous) and typically increases the success rate for useful data. For all experiments the whole-cell configuration was used and three different types of currents were studied; Na+, Ca2+ and K+. Using specific voltage protocols biophysical characteristics of each current was described and compared from single-hole and multi-hole experiments. We showed that currents recorded from these pluripotent stem cell-derived cardiomyocytes are similar to human cardiomyocytes and the response to known pharmacology is as expected. The V0.5 values, I-V relationships, current kinetics and IC50 values determined for known blockers (TTX, nifedipine and cisapride) were comparable for the two recording modes. Clearly the success rate for usable data per measurement plate was significantly increased with the multi-hole technology. This is the first time current properties of stem cell-derived cardiomyocytes have been described from multi-hole recordings with planar automated patch clamp. Our study has shown that automated patch clamp is ready for stem cell-derived exploration.

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