High-Efficient Transfection of Human Embryonic Stem Cells by Single-Cell Plating and Starvation

Stem Cells and Development
Hui LiuXingjun Jiang

Abstract

Nowadays, the low efficiency of small interfering RNA (siRNA) or plasmid DNA (pDNA) transfection is a critical issue in genetic manipulation of human embryonic stem (hES) cells. Development of an efficient transfection method for delivery of siRNAs and plasmids into hES cells becomes more and more imperative. In this study, we tried to modify the traditional transfection protocol by introducing two crucial processes, single-cell plating and starvation, to increase the transfection efficiency in hES cells. Furthermore, we comparatively examined the transfection efficiency of some commercially available siRNA or pDNA transfection reagents in hES cells. Our results showed that the new developed method markedly enhanced the transfection efficiency without influencing the proliferation and pluripotency of hES cells. Lipofectamine RNAiMAX exhibited much higher siRNA transfection efficiency than the other reagents, and FuGENE HD was identified as the best suitable reagent for efficient pDNA transfection of hES cells among the tested reagents.

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Citations

Dec 10, 2019·Biotechnology and Bioengineering·Zhenpeng LiFang Bai
Jan 27, 2019·Stem Cell Research & Therapy·Egle-Helene ErvinToivo Maimets
Jan 23, 2021·Biochemical and Biophysical Research Communications·Kaoru MitsuiKen-Ichiro Kosai

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Methods Mentioned

BETA
gene knockdown
transfection
transfections
PCR
fluorescence microscopy
mechanical dissociation
flow cytometry

Software Mentioned

Bio
SPSS Statistics
Quantity One
Excel
Rad Quantity One

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