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Title 

Improved transfection efficiency and metabolic activity in human embryonic stem cell using non-enzymatic method

Authors 

C Y KimI K HwangC KangE B ChungCho Rok JungH OhY H JeongS H MoonJ S KimK S HongJ H ParkH M Chung

Publisher 

Korean Society for Stem Cell Research

Issue Date 

2018

Citation 

International Journal of Stem Cells

Keywords 

EDTAHuman embryonic stem cellMetabolic activityTransfection

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.

ISSN 

2005-3606

Link 

http://dx.doi.org/10.15283/ijsc18037

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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