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Improved Transfection Efficiency and Metabolic Activity in Human Embryonic Stem Cell Using Non-Enzymatic Method
International Journal of Stem Cells
Published online August 31, 2018;  
© 2018 Korean Society for Stem Cell Research.

C-Yoon Kim1,3,*, In-Kyu Hwang1,*, Changhee Kang1, Eun-Bin Chung1, Cho-Rok Jung2, Hanseul Oh3, Young-Hoon Jeong1, Sung-Hwan Moon1, Jong Soo Kim1, Ki-Sung Hong1, Jae-Hak Park3, Hyung-Min Chung1

1Department of Stem Cell Biology, School of Medicine, Konkuk University, Seoul, Korea
2Gene Therapy Research Unit, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea
3Department of Laboratory Animal Medicine, College of Veterinary Medicine, Seoul National University, Seoul, Korea
Correspondence to: Hyung-Min Chung
Department of Stem Cell Biology, School of Medicine, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Korea
Tel: +82-2-2049-6257, Fax: +82-2-455-9012
E-mail: hmchung@kku.ac.kr
*These authors contributed equally to this work.
Received April 24, 2018; Revised July 12, 2018; Accepted July 12, 2018.
This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Human embryonic stem cells (hESCs) are pluripotent cells widely used in conventional and regenerative medicine due to their ability to self-renew, proliferate and differentiate. Recently, genetic modification of stem cells using genome editing is the most advanced technique for treating hereditary diseases. Nevertheless, the low transfection efficiency of hESCs using enzymatic methods is still limited in in vitro preclinical research. To overcome these limitations, we have developed transfection methods using non-enzymatic treatments on hESCs. In this study, hESCs were transfected following enzymatic (TrypLE and trypsin) and non-enzymatic treatment ethylenediaminetetraacetic acid (EDTA) to increase transfection efficiency. Flow cytometric analysis using an enhanced green fluorescent protein vector showed a significantly increased transfection efficiency of EDTA method compared to standard enzyme method. In addition, the EDTA approach maintained stable cell viability and recovery rate of hESCs after transfection. Also, metabolic activity by using Extracellular Flux Analyzer revealed that EDTA method maintained as similar levels of cell functionality as normal group comparing with enzymatic groups. These results suggest that transfection using EDTA is a more efficient and safe substitute for transfection than the use of standard enzymatic methods.
Keywords : Human embryonic stem cell, Transfection, EDTA, Metabolic activity