Controllable Large-Scale Transfection of Primary Mammalian Cardiomyocytes on a Nanochannel Array Platform.
Controllable Large-Scale Transfection of Primary Mammalian Cardiomyocytes on a Nanochannel Array Platform.
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DOI：
10.1002/smll.201601465
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发表时间：
2016-11
期刊：
影响因子：
13.3
通讯作者：
Lee, Ly James
中科院分区：
文献类型：
--
作者：
Chang, Lingqian;Gallego-Perez, Daniel;Chiang, Chi-Ling;Bertani, Paul;Kuang, Tairong;Sheng, Yan;Chen, Feng;Chen, Zhou;Shi, Junfeng;Yang, Hao;Huang, Xiaomeng;Malkoc, Veysi;Lu, Wu;Lee, Ly James
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. An nanochannel array platform trap and precisely transfect primary adult mammalian cardiomyocytes with high efficiency /cell viability. A simple ‘dewetting‘ approach precisely positions single cell on chip. Nanochannel-based electroporation can determine the dose of delivery, offering quality control of microRNA-based gene therapy in cardiovascular cells as demonstrated by delivering miR-29 into primary cadiomyocytes for determining dose effect for apoptosis.
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影响因子：
7.4
作者：
Henslee, Brian E.;Morss, Andrew;Lee, L. James
通讯作者：
Lee, L. James
影响因子：
6.1
作者：
Kang W;Giraldo-Vela JP;Nathamgari SS;McGuire T;McNaughton RL;Kessler JA;Espinosa HD
通讯作者：
Espinosa HD
影响因子：
13.3
作者：
Gao, Keliang;Li, Lei;Lee, L. James
通讯作者：
Lee, L. James
影响因子：
6.1
作者：
Khine, M;Lau, A;Lee, LP
通讯作者：
Lee, LP
DOI：
10.1158/1078-0432.ccr-12-3191
发表时间：
2013-05-01
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Huang X;Schwind S;Yu B;Santhanam R;Wang H;Hoellerbauer P;Mims A;Klisovic R;Walker AR;Chan KK;Blum W;Perrotti D;Byrd JC;Bloomfield CD;Caligiuri MA;Lee RJ;Garzon R;Muthusamy N;Lee LJ;Marcucci G
通讯作者：
Marcucci G