Effective naked plasmid DNA delivery into stem cells by microextrusion-based transient-transfection system for in situ cardiac repair
| Autor: | Shan-hui Hsu, Chii-Ming Lee, Nien-Chi Huang |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Cancer Research Immunology Polyurethanes Cell- and Tissue-Based Therapy Microextrusion Gene delivery Transfection Umbilical Cord 03 medical and health sciences 0302 clinical medicine Plasmid medicine Immunology and Allergy Animals Myocytes Cardiac Cell damage Genetics (clinical) Zebrafish Transplantation Chemistry fungi Mesenchymal stem cell Heart Hydrogels Mesenchymal Stem Cells Cell Biology DNA Genetic Therapy medicine.disease Cellular Reprogramming Cell biology GATA4 Transcription Factor 030104 developmental biology Oncology 030220 oncology & carcinogenesis embryonic structures Self-healing hydrogels Stem cell Plasmids |
| Zdroj: | Cytotherapy. 22(2) |
| ISSN: | 1477-2566 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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