Highly efficient transfection of human induced pluripotent stem cells using magnetic nanoparticles

Megan A. Yamoah, Maryam Moshref, Janhavi Sharma, Wei Chun Chen, Hannah A. Ledford, Jeong Han Lee, Karen S. Chavez, Wenying Wang, Javier E Lopez, Deborah Lieu, Padmini Sirish, Xiao Dong Zhang

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)6073-6078
Number of pages6
JournalInternational Journal of Nanomedicine
Volume13
DOIs
StatePublished - Jan 1 2018

Fingerprint

Induced Pluripotent Stem Cells
Stem cells
Nanoparticles
Transfection
Nucleic acids
Nucleic Acids
HEK293 Cells
Genes
Flow cytometry
Confocal microscopy
Clamping devices
Magnetic Fields
Cell membranes
Cytotoxicity
Transgenes
Cardiac Myocytes
Confocal Microscopy
Regeneration
Flow Cytometry
Cell Membrane

Keywords

  • efficiency
  • human induced pluripotent stem cell-derived cardiomyocytes
  • pluripotency
  • therapy

ASJC Scopus subject areas

  • Biophysics
  • Bioengineering
  • Biomaterials
  • Pharmaceutical Science
  • Drug Discovery
  • Organic Chemistry

Cite this

Yamoah, M. A., Moshref, M., Sharma, J., Chen, W. C., Ledford, H. A., Lee, J. H., ... Zhang, X. D. (2018). Highly efficient transfection of human induced pluripotent stem cells using magnetic nanoparticles. International Journal of Nanomedicine, 13, 6073-6078. https://doi.org/10.2147/IJN.S172254

Highly efficient transfection of human induced pluripotent stem cells using magnetic nanoparticles. / Yamoah, Megan A.; Moshref, Maryam; Sharma, Janhavi; Chen, Wei Chun; Ledford, Hannah A.; Lee, Jeong Han; Chavez, Karen S.; Wang, Wenying; Lopez, Javier E; Lieu, Deborah; Sirish, Padmini; Zhang, Xiao Dong.

In: International Journal of Nanomedicine, Vol. 13, 01.01.2018, p. 6073-6078.

Research output: Contribution to journalArticle

Yamoah, MA, Moshref, M, Sharma, J, Chen, WC, Ledford, HA, Lee, JH, Chavez, KS, Wang, W, Lopez, JE, Lieu, D, Sirish, P & Zhang, XD 2018, 'Highly efficient transfection of human induced pluripotent stem cells using magnetic nanoparticles', International Journal of Nanomedicine, vol. 13, pp. 6073-6078. https://doi.org/10.2147/IJN.S172254
Yamoah, Megan A. ; Moshref, Maryam ; Sharma, Janhavi ; Chen, Wei Chun ; Ledford, Hannah A. ; Lee, Jeong Han ; Chavez, Karen S. ; Wang, Wenying ; Lopez, Javier E ; Lieu, Deborah ; Sirish, Padmini ; Zhang, Xiao Dong. / Highly efficient transfection of human induced pluripotent stem cells using magnetic nanoparticles. In: International Journal of Nanomedicine. 2018 ; Vol. 13. pp. 6073-6078.
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abstract = "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.",
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AU - Yamoah, Megan A.

AU - Moshref, Maryam

AU - Sharma, Janhavi

AU - Chen, Wei Chun

AU - Ledford, Hannah A.

AU - Lee, Jeong Han

AU - Chavez, Karen S.

AU - Wang, Wenying

AU - Lopez, Javier E

AU - Lieu, Deborah

AU - Sirish, Padmini

AU - Zhang, Xiao Dong

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N2 - 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.

AB - 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.

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KW - pluripotency

KW - therapy

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