title: Evaluation of Changes in Morphology and Function of Human Induced Pluripotent Stem Cell Derived Cardiomyocytes (HiPSC-CMs) Cultured on an Aligned-Nanofiber Cardiac Patch.
abstract: BACKGROUND
  Dilated cardiomyopathy is a major cause of progressive heart failure. Utilization of stem cell therapy offers a potential means of regenerating viable cardiac tissue. However, a major obstacle to stem cell therapy is the delivery and survival of implanted stem cells in the ischemic heart. To address this issue, we have developed a biomimetic aligned nanofibrous cardiac patch and characterized the alignment and function of human inducible pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) cultured on this cardiac patch. This hiPSC-CMs seeded patch was compared with hiPSC-CMs cultured on standard flat cell culture plates.
  
  
  METHODS
  hiPSC-CMs were cultured on; 1) a highly aligned polylactide-co-glycolide (PLGA) nanofiber scaffold (~50 microns thick) and 2) on a standard flat culture plate. Scanning electron microscopy (SEM) was used to determine alignment of PLGA nanofibers and orientation of the cells on the respective surfaces. Analysis of gap junctions (Connexin-43) was performed by confocal imaging in both the groups. Calcium cycling and patch-clamp technique were performed to measure calcium transients and electrical coupling properties of cardiomyocytes.
  
  
  RESULTS
  SEM demonstrated >90% alignment of the nanofibers in the patch which is similar to the extracellular matrix of decellularized rat myocardium. Confocal imaging of the cardiomyocytes demonstrated symmetrical alignment in the same direction on the aligned nanofiber patch in sharp contrast to the random appearance of cardiomyocytes cultured on a tissue culture plate. The hiPSC-CMs cultured on aligned nanofiber cardiac patches showed more efficient calcium cycling compared with cells cultured on standard flat surface culture plates. Quantification of mRNA with qRT-PCR confirmed that these cardiomyocytes expressed α-actinin, troponin-T and connexin-43 in-vitro.
  
  
  CONCLUSIONS
  Overall, our results demonstrated changes in morphology and function of human induced pluripotent derived cardiomyocytes cultured in an anisotropic environment created by an aligned nanofiber patch. In this environment, these cells better approximate normal cardiac tissue compared with cells cultured on flat surface and can serve as the basis for bioengineering of an implantable cardiac patch.
authors.affiliation: Department of Emergency Medicine, Davis Heart Lung Research Institute, The Ohio State University Wexner Medical Center, Columbus, OH, United States of America.
authors.forename: Mahmood
authors.initials: M
authors.lastname: Khan
authors.affiliation: Department of Materials Science and Engineering, Davis Heart Lung Research Institute, Ohio State University, Columbus, OH, United States of America.
authors.forename: Yanyi
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Department of Emergency Medicine, Davis Heart Lung Research Institute, The Ohio State University Wexner Medical Center, Columbus, OH, United States of America.
authors.forename: Serena
authors.initials: S
authors.lastname: Hua
authors.affiliation: Nanofiber Solutions, Columbus, OH, United States of America.
authors.forename: Jed
authors.initials: J
authors.lastname: Johnson
authors.affiliation: Department of Physiology and Cell Biology, Davis Heart Lung Research Institute, The Ohio State University Wexner Medical Center, Columbus, OH, United States of America.
authors.forename: Andriy
authors.initials: A
authors.lastname: Belevych
authors.affiliation: Department of Physiology and Cell Biology, Davis Heart Lung Research Institute, The Ohio State University Wexner Medical Center, Columbus, OH, United States of America.
authors.forename: Paul M L
authors.initials: PM
authors.lastname: Janssen
authors.affiliation: Department of Physiology and Cell Biology, Davis Heart Lung Research Institute, The Ohio State University Wexner Medical Center, Columbus, OH, United States of America.
authors.forename: Sandor
authors.initials: S
authors.lastname: Gyorke
authors.affiliation: Department of Materials Science and Engineering, Davis Heart Lung Research Institute, Ohio State University, Columbus, OH, United States of America.
authors.forename: Jianjun
authors.initials: J
authors.lastname: Guan
authors.affiliation: Department of Emergency Medicine, Davis Heart Lung Research Institute, The Ohio State University Wexner Medical Center, Columbus, OH, United States of America.
authors.forename: Mark G
authors.initials: MG
authors.lastname: Angelos
mesh_terms: D000200:Action Potentials; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism; D057139:Nanofibers; D054457:Tissue Scaffolds
chemical_list: D002118:Calcium