Highly efficient transfection of human induced pluripotent stem cells using magnetic nanoparticles
Megan A Yamoah,1 Maryam Moshref,1 Janhavi Sharma,1 Wei Chun Chen,1 Hannah A Ledford,1 Jeong Han Lee,2 Karen S Chavez,1 Wenying Wang,2 Javier E López,1 Deborah K Lieu,1 Padmini Sirish,1,3 Xiao-Dong Zhang1,3 1Department of Internal Medicine, University of California, Davis, CA, USA; 2Depar...
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| Autores principales: | , , , , , , , , , , , |
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| Formato: | article |
| Lenguaje: | EN |
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Dove Medical Press
2018
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| Materias: | |
| Acceso en línea: | https://doaj.org/article/b521378c08d14e7599813c9375db2781 |
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| Sumario: | Megan A Yamoah,1 Maryam Moshref,1 Janhavi Sharma,1 Wei Chun Chen,1 Hannah A Ledford,1 Jeong Han Lee,2 Karen S Chavez,1 Wenying Wang,2 Javier E López,1 Deborah K Lieu,1 Padmini Sirish,1,3 Xiao-Dong Zhang1,3 1Department of Internal Medicine, University of California, Davis, CA, USA; 2Department of Physiology and Cell Biology, University of Nevada, Reno, NV, USA; 3Department of Veterans Affairs, Northern California Health Care System, Mather, CA, USA Purpose: The delivery of transgenes into human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (hiPSC-CMs) represents an important tool in cardiac regeneration with potential for clinical applications. Gene transfection is more difficult, however, for hiPSCs and hiPSC-CMs than for somatic cells. Despite improvements in transfection and transduction, the efficiency, cytotoxicity, safety, and cost of these methods remain unsatisfactory. The objective of this study is to examine gene transfection in hiPSCs and hiPSC-CMs using magnetic nanoparticles (NPs).Methods: Magnetic NPs are unique transfection reagents that form complexes with nucleic acids by ionic interaction. The particles, loaded with nucleic acids, can be guided by a magnetic field to allow their concentration onto the surface of the cell membrane. Subsequent uptake of the loaded particles by the cells allows for high efficiency transfection of the cells with nucleic acids. We developed a new method using magnetic NPs to transfect hiPSCs and hiPSC-CMs. HiPSCs and hiPSC-CMs were cultured and analyzed using confocal microscopy, flow cytometry, and patch clamp recordings to quantify the transfection efficiency and cellular function.Results: We compared the transfection efficiency of hiPSCs with that of human embryonic kidney (HEK 293) cells. We observed that the average efficiency in hiPSCs was 43%±2% compared to 62%±4% in HEK 293 cells. Further analysis of the transfected hiPSCs showed that the differentiation of hiPSCs to hiPSC-CMs was not altered by NPs. Finally, robust transfection of hiPSC-CMs with an efficiency of 18%±2% was obtained.Conclusion: The difficult-to-transfect hiPSCs and hiPSC-CMs were efficiently transfected using magnetic NPs. Our study offers a novel approach for transfection of hiPSCs and hiPSC-CMs without the need for viral vector generation. Keywords: human induced pluripotent stem cell-derived cardiomyocytes, therapy, pluripotency, efficiency |
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