Effective naked plasmid DNA delivery into stem cells by microextrusion-based transient-transfection system for in situ cardiac repair

被引:10
作者
Huang, Nien-Chi [1 ]
Lee, Chii-Ming [2 ]
Hsu, Shan-hui [1 ,3 ,4 ]
机构
[1] Natl Taiwan Univ, Inst Polymer Sci & Engn, 1,Sec 4,Roosevelt Rd, Taipei 10617, Taiwan
[2] Natl Taiwan Univ Hosp, Dept Internal Med, Div Cardiol, Taipei, Taiwan
[3] Natl Taiwan Univ, Ctr Tissue Engn & 3D Printing, Taipei, Taiwan
[4] Natl Hlth Res Inst, Inst Cellular & Syst Med, Zhunan, Taiwan
关键词
cardiac repair; cell reprogramming; in situ transfection; naked plasmid delivery; polyurethane hydrogels; NONVIRAL GENE DELIVERY; POLYURETHANE ELASTOMER; HEART REGENERATION; HUMAN FIBROBLASTS; HYDROGELS; TOXICITY; CARDIOMYOCYTES; BIOMATERIALS; CONDUCTION; POLYPLEXES;
D O I
10.1016/j.jcyt.2019.12.003
中图分类号
Q813 [细胞工程];
学科分类号
摘要
Background aims. Combining the use of transfection reagents and physical methods can markedly improve the efficiency of gene delivery; however, such methods often cause cell damage. Additionally, naked plasmids without any vector or physical stimulation are difficult to deliver into stem cells. In this study, we demonstrate a simple and rapid method to simultaneously facilitate efficient in situ naked gene delivery and form a bioactive hydrogel scaffold. Methods. Transfecting naked GATA binding protein 4 (GATA4) plasmids into human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) by co-extruding naked plasmids and hUC-MSCs with a biomimetic and negatively charged water-based biodegradable thermo-responsive polyurethane (PU) hydrogel through a microextrusion-based transient-transfection system can upregulate the other cardiac marker genes. Results. The PU hydrogels with optimized physicochemical properties (such as hard-soft segment composition, size, hardness and thermal gelation) induced GATA4-transfected hUC-MSCs to express the cardiac marker proteins and then differentiated into cardiomyocyte-like cells in 15 days. We further demonstrated that GATA4-transfected hUC-MSCs in PU hydrogel were capable of in situ revival of heart function in zebrafish in 30 days. Conclusions. Our results suggest that hUC-MSCs and naked plasmids encapsulated in PU hydrogels might represent a new strategy for in situ tissue therapy using the microextrusion-based transient-transfection system described here. This transfection system is simple, effective and safer than conventional technologies. (C) 2019 International Society for Cell and Gene Therapy. Published by Elsevier Inc. All rights reserved.
引用
收藏
页码:70 / 81
页数:12
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