title: Methods for assessing the electromechanical integration of human pluripotent stem cell-derived cardiomyocyte grafts.
abstract: Cardiomyocytes derived from human pluripotent stem cells show tremendous promise for the replacement of myocardium and contractile function lost to infarction. However, until recently, no methods were available to directly determine whether these stem cell-derived grafts actually couple with host myocardium and fire synchronously following transplantation in either intact or injured hearts. To resolve this uncertainty, our group has developed techniques for the intravital imaging of hearts engrafted with stem cell-derived cardiomyocytes that have been modified to express the genetically encoded protein calcium sensor, GCaMP. When combined with the simultaneously recorded electrocardiogram, this protocol allows one to make quantitative assessments as to the presence and extent of host-graft electrical coupling as well as the timing and pattern of graft activation. As described here, this system has been employed to investigate the electromechanical integration of human embryonic stem cell-derived cardiomyocytes in a guinea pig model of cardiac injury, but analogous approaches should be applicable to other human graft cell types and animal models.
authors: Department of Pathology, Institute for Stem Cells and Regenerative Medicine, University of Washington, Seattle, WA, 98109, USA.
authors: Wei-Zhong
authors: WZ
authors: Zhu
authors: Dominic
authors: D
authors: Filice
authors: Nathan J
authors: NJ
authors: Palpant
authors: Michael A
authors: MA
authors: Laflamme
mesh_terms: D000818:Animals; D002460:Cell Line; D015925:Cryopreservation; D003851:Deoxyribonucleases / Q000737:chemistry / Q000378:metabolism; D018274:Electroporation; D006168:Guinea Pigs; D006335:Heart Injuries / Q000473:pathology; D006801:Humans; D008297:Male; D055595:Mechanical Phenomena; D057054:Molecular Imaging; D032383:Myocytes, Cardiac / Q000166:cytology*; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation*; D033581:Stem Cell Transplantation / Q000379:methods*; D016335:Zinc Fingers
chemical_list: D003851:Deoxyribonucleases