title: Controllable Large-Scale Transfection of Primary Mammalian Cardiomyocytes on a Nanochannel Array Platform.
abstract: While electroporation has been widely used as a physical method for gene transfection in vitro and in vivo, its application in gene therapy of cardiovascular cells remains challenging. Due to the high concentration of ion-transport proteins in the sarcolemma, conventional electroporation of primary cardiomyocytes tends to cause ion-channel activation and abnormal ion flux, resulting in low transfection efficiency and high mortality. In this work, a high-throughput nanoelectroporation technique based on a nanochannel array platform is reported, which enables massively parallel delivery of genetic cargo (microRNA, plasmids) into mouse primary cardiomyocytes in a controllable, highly efficient, and benign manner. A simple "dipping-trap" approach was implemented to precisely position a large number of cells on the nanoelectroporation platform. With dosage control, our device precisely titrates the level of miR-29, a potential therapeutic agent for cardiac fibrosis, and determines the minimum concentration of miR-29 causing side effects in mouse primary cardiomyocytes. Moreover, the dose-dependent effect of miR-29 on mitochondrial potential and homeostasis is monitored. Altogether, our nanochannel array platform provides efficient trapping and transfection of primary mouse cardiomyocyte, which can improve the quality control for future microRNA therapy in heart diseases.
authors.affiliation: NSEC Center for Affordable Nanoengineering of Polymeric Biomedical Devices, Ohio State University, Columbus, OH, 43210, USA.|Department of Biomedical Engineering, Ohio State University, Columbus, OH, 43209, USA.
authors.forename: Lingqian
authors.initials: L
authors.lastname: Chang
authors.affiliation: NSEC Center for Affordable Nanoengineering of Polymeric Biomedical Devices, Ohio State University, Columbus, OH, 43210, USA.|Department of Biomedical Engineering, Ohio State University, Columbus, OH, 43209, USA.|Department of Surgery, Center for Regenerative Medicine and Cell-based Therapies, Ohio State University, Columbus, OH, 43209, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Gallego-Perez
authors.affiliation: NSEC Center for Affordable Nanoengineering of Polymeric Biomedical Devices, Ohio State University, Columbus, OH, 43210, USA.|Department of Internal Medicine, The Ohio State University, Columbus, OH, 43209, USA.
authors.forename: Chi-Ling
authors.initials: CL
authors.lastname: Chiang
authors.affiliation: Electrical and Computer Engineering Department, Ohio State University, Columbus, OH, 43209, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Bertani
authors.affiliation: NSEC Center for Affordable Nanoengineering of Polymeric Biomedical Devices, Ohio State University, Columbus, OH, 43210, USA.
authors.forename: Tairong
authors.initials: T
authors.lastname: Kuang
authors.affiliation: NSEC Center for Affordable Nanoengineering of Polymeric Biomedical Devices, Ohio State University, Columbus, OH, 43210, USA.|Department of Chemical and Biomolecular Engineering, Ohio State University, Columbus, OH, 43210, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Sheng
authors.affiliation: NSEC Center for Affordable Nanoengineering of Polymeric Biomedical Devices, Ohio State University, Columbus, OH, 43210, USA.|Department of Chemical and Biomolecular Engineering, Ohio State University, Columbus, OH, 43210, USA.
authors.forename: Feng
authors.initials: F
authors.lastname: Chen
authors.affiliation: NSEC Center for Affordable Nanoengineering of Polymeric Biomedical Devices, Ohio State University, Columbus, OH, 43210, USA.|College of Mechanical and Power Engineering, Nanjing Tech University, Nanjing, 211800, China.
authors.forename: Zhou
authors.initials: Z
authors.lastname: Chen
authors.affiliation: NSEC Center for Affordable Nanoengineering of Polymeric Biomedical Devices, Ohio State University, Columbus, OH, 43210, USA.
authors.forename: Junfeng
authors.initials: J
authors.lastname: Shi
authors.affiliation: Electrical and Computer Engineering Department, Ohio State University, Columbus, OH, 43209, USA.
authors.forename: Hao
authors.initials: H
authors.lastname: Yang
authors.affiliation: NSEC Center for Affordable Nanoengineering of Polymeric Biomedical Devices, Ohio State University, Columbus, OH, 43210, USA.|Department of Internal Medicine, The Ohio State University, Columbus, OH, 43209, USA.
authors.forename: Xiaomeng
authors.initials: X
authors.lastname: Huang
authors.affiliation: NSEC Center for Affordable Nanoengineering of Polymeric Biomedical Devices, Ohio State University, Columbus, OH, 43210, USA.|Department of Chemical and Biomolecular Engineering, Ohio State University, Columbus, OH, 43210, USA.
authors.forename: Veysi
authors.initials: V
authors.lastname: Malkoc
authors.affiliation: NSEC Center for Affordable Nanoengineering of Polymeric Biomedical Devices, Ohio State University, Columbus, OH, 43210, USA.|Electrical and Computer Engineering Department, Ohio State University, Columbus, OH, 43209, USA.
authors.forename: Wu
authors.initials: W
authors.lastname: Lu
authors.affiliation: NSEC Center for Affordable Nanoengineering of Polymeric Biomedical Devices, Ohio State University, Columbus, OH, 43210, USA.|Department of Biomedical Engineering, Ohio State University, Columbus, OH, 43209, USA.|Chemical and Biomolecular Engineering Department, Ohio State University, Columbus, OH, 43209, USA.|Department of Chemical and Biomolecular Engineering, Ohio State University, Columbus, OH, 43210, USA.
authors.forename: Ly James
authors.initials: LJ
authors.lastname: Lee
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003198:Computer Simulation; D018274:Electroporation; D051379:Mice; D035683:MicroRNAs / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000378:metabolism*; D053758:Nanoparticles / Q000737:chemistry*; D036103:Nanotechnology / Q000379:methods*; D014162:Transfection / Q000379:methods*
chemical_list: C547014:MIRN29 microRNA, mouse; D035683:MicroRNAs
keywords: miR29; nanochannel arrays; nanoelectroporation; nanofabrication; primary cardiomyocytes