International Journal of Stem Cells

  • 제16권1호
  • /
  • Pages.117-122
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  • 2023
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  • 2005-3606(pISSN)
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  • 2005-5447(eISSN)
한국줄기세포학회 (Korean Society for Stem Cell Research)
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Reduced Cytotoxicity by Repetitive mRNA Transfection in Differentiated Neurons

  • Seung Hwan Ko (Graduate School of Biomedical Science and Engineering, Hanyang University) ;
  • Jin Sun Kang (Graduate School of Biomedical Science and Engineering, Hanyang University) ;
  • Sang-Mi Kim (Hanyang Biomedical Research Institute, Hanyang University) ;
  • Eun-Hye Lee (Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, Johns Hopkins University School of Medicine) ;
  • Chang-Hwan Park (Graduate School of Biomedical Science and Engineering, Hanyang University)
  • 투고 : 2022.07.19
  • 심사 : 2022.11.07
  • 발행 : 2023.02.28
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초록

Background and Objectives: mRNA-based protein expression technology has been used to express functional proteins. We have previously generated dopamine neurons from rat-embryo derived neural precursor cells (NPCs) through repeated transfection of synthetic transcription factor mRNA encoding dopamine-inducible genes. However, NPCs began to die approximately 10 d post-transfection. In this study, we examined a long-term transfection protocol that did not affect cell viability. Methods and Results: Experiments were performed in eight groups sorted according to the start date of mRNA transfection. mRNA was transfected into NPCs daily for 21 d and live cell images of each group were recorded. NPCs which were differentiated for more than five days showed sustained gene expression and appreciable viability despite daily mRNA transfection for 21 d. Conclusions: Repeated mRNA transfection requires cells with a sufficient differentiation period.

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과제정보

This work was supported by the Individual Basic Science and Engineering Research Program (2019R1A2C2005681) of the National Research Foundation of the Ministry of Science and ICT in Korea.

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