M Kolahdouz Mohammadi - Department of Biology, School of Basic Science, Science andResearch Branch, Islamic Azad University, Tehran, Iran . Department of S tem Cells and Developmental Biology, Cell ScienceResearch Center, Royan Ins titute for S tem Cell Biology andTechnology, AC
S Pahlavan - Department of S tem Cells and Developmental Biology, Cell ScienceResearch Center, Royan Ins titute for S tem Cell Biology andTechnology, ACECR, Tehran, Iran
N Salehi - School of Biological Science, Ins titute for Research in FundamentalSciences (IPM), Tehran, Iran
F SotoodehnejadNematalah - Department of Biology, School of Basic Science, Science andResearch Branch, Islamic Azad University, Tehran, Iran
M Totonchi - Department of S tem Cells and Developmental Biology, Cell ScienceResearch Center, Royan Ins titute for S tem Cell Biology andTechnology, ACECR, Tehran, Iran

Objective: It has been shown that autophagy, an essentialcatabolic process for survival under s tress, contributes to differentiationinto various types of cells such as cardiomyocytes.Adenosine ۵ʹ-monophosphate (AMP)-activated protein kinase(AMPK) is an energy-sensing protein kinase involved in the regulation of autophagy. It has been demons trated that AMPKplays a role in many other cellular processes. Due to its involvementin multiple cellular processes, AMPK can influence thesurvival and health of cardiomyocytes. In this s tudy, the timedependenteffects of an activator of AMPK (Metformin) and anautophagy inhibitor (Hydroxychloroquine) on human pluripotents tem cell-derived cardiomyocytes (hPSC-CM) differentiationwere inves tigated.Materials and Methods: During cardiomyocyte differentiation,Metformin and Hydroxychloroquine were used to induceand inhibit autophagy, respectively. Days ۰, ۲, ۴, and ۱۳ wereexamined for the expression patterns of the autophagy genes.Results: During cardiomyocyte differentiation, autophagy wasupregulated, and it was not affected by autophagy inhibitor/inducer.AMPK activation also increased the expression of cardiomyocyte-specific markers in hPSC-CMs. Also, autophagy inhibitionby targeting autophagosome-lysosome fusion impairedcardiomyocyte differentiation.Conclusion: In conclusion, AMPK might be a promising targetfor regulating cardiomyocyte generation by in vitro differentiationof pluripotent s tem cells, due to the importance of autophagyin this cellular process.

Autophagy, Adenosine ۵ʹ-monophosphate-activatedprotein kinase (AMPK), Cardiomyocyte Differentiation, HumanPluripotent S tem Cell (hPSC)