Dual optical recordings for action potentials and calcium handling in induced pluripotent stem cell models of cardiac arrhythmias using genetically encoded fluorescent indicators

Stem Cells Translational Medicine
LouJin SongMasayuki Yazawa

Abstract

Reprogramming of human somatic cells to pluripotency has been used to investigate disease mechanisms and to identify potential therapeutics. However, the methods used for reprogramming, in vitro differentiation, and phenotyping are still complicated, expensive, and time-consuming. To address the limitations, we first optimized a protocol for reprogramming of human fibroblasts and keratinocytes into pluripotency using single lipofection and the episomal vectors in a 24-well plate format. This method allowed us to generate multiple lines of integration-free and feeder-free induced pluripotent stem cells (iPSCs) from seven patients with cardiac diseases and three controls. Second, we differentiated human iPSCs derived from patients with Timothy syndrome into cardiomyocytes using a monolayer differentiation method. We found that Timothy syndrome cardiomyocytes showed slower, irregular contractions and abnormal calcium handling compared with the controls. The results are consistent with previous reports using a retroviral method for reprogramming and an embryoid body-based method for cardiac differentiation. Third, we developed an efficient approach for recording the action potentials and calcium transients simultaneously in control...Continue Reading

References

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Citations

Dec 24, 2015·Frontiers in Cell and Developmental Biology·Nazanin F DolatshadGabor Földes
Oct 30, 2016·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·Matthew C WhiteXi Yang
Sep 20, 2016·British Journal of Pharmacology·Luca SalaChristine L Mummery
Jun 27, 2017·Stem Cell Reports·LouJin SongMasayuki Yazawa
Sep 9, 2017·Disease Models & Mechanisms·Karina O BrandãoRichard P Davis
Nov 15, 2019·Cell Transplantation·Aline Yen Ling Wang, Charles Yuen Yung Loh
Feb 22, 2017·Current Cardiology Reports·Alexander GoedelAlessandra Moretti
Jul 12, 2020·Nature Communications·Nao NittaKeisuke Goda
Apr 17, 2020·Frontiers in Cardiovascular Medicine·Pim R R van GorpAntoine A F de Vries
Jul 22, 2019·Experimental Biology and Medicine·Dan HanGuoliang Li
Dec 20, 2020·Nature Reviews. Cardiology·Emilia Entcheva, Matthew W Kay
Apr 4, 2021·International Journal of Molecular Sciences·Aleksandra NijakMaaike Alaerts
Oct 27, 2021·The Journal of General Physiology·Marina AngeliniRiccardo Olcese

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