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, we report a high-throughput nano-electroporation technique based on a nanochannel array platform, which enables massively parallel delivery of genetic cargo (microRNA, plasmids) into mouse primary cardiomyocytes in a controllable, highly efficient and benign manner. A simple ‘dewetting’ approach was implemented to precisely position a large number of cells on the nano-electroporation platform. With dosage control, our device precisely titrated the level of miR-29, a potential therapeutic agent for cardiac fibrosis, and determined 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 was monitored. Altogether, our nanochannel array platform provides efficient trapping and transfection of primary mouse cardiomyocyte, which could improve the quality control for future microRNA therapy in heart diseases.

Multifurcate Assembly of Slanted Micropillars Fabricated by Superposition of Optical Vortices and Application in High‐Efficiency Trapping Microparticles

“…The ability of manipulating particles desired in biological devices and chemical analysis has drawn crucial attentions from researchers . For example, substantial mechanical grippers are used to capture, hold, and analyze living cells for genomics, proteomics, and diagnostics .…”

“…According to the previous studies, a variety of methods, such as microlens arrays, spatial light modulator (SLM), and multibeam interference with four‐titled plane waves have been developed for rapid laser fabrication, but these methods have not yet been explored for fabricating furcate pillars. In addition, the assembled micropillars have received much attention recently because their ability in trapping microobjects is potentially used in the fields of biomedical devices and chemical analysis . However, one challenge of these applications is the relatively low trapping ratio (<40%) because only a part of the straight micro/nanopillars which are used as grippers can catch particles successfully when capillary force driving the pillars to self‐assembly.…”

Light‐Responsive Polymer Membranes

“…Applications of MSMs as smart sensors and gated switches use MCM‐41 and SBA‐15 particles functionalized with responsive polymer brushes, able to control the pore opening and closing in response to the surrounding physicochemical changes.…”

Wearable Devices for Single-Cell Sensing and Transfection

“…Despite its feasibility, the application of bulk electroporation in vivo remains limited, because a large electric potential required to generate sufficient transmembrane potential leads to significant cell death and low efficiency [72]. By contrast, singlecell electroporation shows superior performance in terms of specificity, dosage control, cell viability, and transfection efficiency [64,76]. In single-cell electroporation, a nanochannel connected to a microfluidic reservoir and filled with electrolytes deploys the electrical potential to a small patch of the cell membrane.…”