Abstract
With recent advances in stem cell technology, it is becoming efficient to differentiate human pluripotent stem cells (hPSCs) into cardiomyocytes, which can subsequently be used for myriad purposes, ranging from interrogating mechanisms of cardiovascular disease, developing novel cellular therapeutic approaches, as well as assessing the cardiac safety profile of compounds. However, the relative inability to acquire abundant pure and mature cardiomyocytes still hinders these applications. Recently, it was reported that glucose-depleted culture medium supplemented with lactate can facilitate purification of hPSC-derived cardiomyocytes. Here, we report that fatty acid as a lactate replacement has not only a similar purification effect but also improves the electrophysiological characteristics of hPSC-derived cardiomyocytes. Glucose-depleted culture medium supplemented with fatty acid and 3,3′,5-Triiodo-l-thyronine (T3) was used during enrichment of hPSC-derived cardiomyocytes. Compared to untreated control cells, the treated cardiomyocytes exhibited enhanced action potential (AP) maximum upstroke velocity (as shown by a significant increase in dV/dtmax), action potential amplitude, as well as AP duration at 50% (APD50) and 90% (APD90) of repolarization. The treated cardiomyocytes displayed higher sensitivity to isoproterenol, more organized sarcomeric structures, and lower proliferative activity. Expression profiling showed that various ion channel and cardiac-specific genes were elevated as well. Our results suggest that the use of fatty acid and T3 can facilitate purification and maturation of hPSC-derived cardiomyocytes.
Highlights
Recent developments in stem cell technology hold promise for use in clinical applications
We found that glucose-depleted culture medium supplemented with fatty acid and T3 can be used for purification of human pluripotent stem cells (hPSCs)-derived cardiomyocytes
HL-1 cells were cultured in non-glucose medium, non-glucose medium supplemented with lactate, and non-glucose medium supplemented with fatty acid (Figure S1 in Supplementary Material)
Summary
Recent developments in stem cell technology hold promise for use in clinical applications. New differentiation methods enable rapid and efficient generation of human pluripotent stem cell (hPSC)-derived cardiomyocytes [1, 2], which can subsequently be utilized as a powerful platform for cardiovascular disease modeling and moderate- to high-throughput. Several methods have been used for cardiomyocyte purification [4], including percoll gradient centrifugation [5], genetic modification [to generate expression of an identifier molecule driven by a cardiac-specific promoter [6, 7]], and mitochondrial dyes or antibodies against cardiac specific markers [8,9,10,11] These techniques all possess significant drawbacks, including low efficiency (percoll gradient centrifugation), safety concerns for therapeutic applications (genetic modification), and relatively high experimental cost (mitochondrial dyes and cardiac specific markers). In the first report of cardiomyocyte purification by using a metabolic method, Fukuda and colleagues demonstrated that glucose-depleted culture medium supplemented with lactate could be used to purify mouse and human PSC-derived cardiomyocytes [12]
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
