Optimal conditions for transfecting Wharton’s Jelly-derived mesenchymal stem cells with electroporation method

doi:10.3969/j.issn.2095-4344.0870

Abstract

Abstract:

BACKGROUND: Wharton’s Jelly-derived mesenchymal stem cells are relatively primitive stem cells that are ideal vectors for gene therapy. However, there is a lack of studies on the conditions for the electrotransfection of Wharton’s Jelly-derived mesenchymal stem cells. Therefore, exploring the optimal conditions for the electrotransfection of Wharton’s Jelly-derived mesenchymal stem cells occupies an important position.
OBJECTIVE: To investigate the effects of different electroporation conditions on the transfection efficiency of Wharton’s Jelly-derived mesenchymal stem cells, and to explore the optimal conditions for cell electroporation.
METHODS: By controlling the transfection conditions such as voltage, pulse duration and cell status, EEV-EGFP plasmids were transfected into Wharton’s Jelly-derived mesenchymal stem cells by electroporation under different conditions. Transfection efficiency was detected by flow cytometry.
RESULTS AND CONCLUSION: (1) The transfection efficiency was intended to increase when the voltage ranged from 125 V to 150 V, and the maximum transfection efficiency was obtained when the voltage was 150 V. However, when the voltage was further increased to 170 V, the transfection efficiency began to decrease considerably. (2) The maximum transfection efficiency was obtained when the pulse duration was 5.0 ms, while it was certainly decreased when the pulse duration was 2.5 and 7.5 ms. (3) The transfection efficiency of the cells cultured under normoxia was higher than that under hypoxic culture. These findings reveal that normally cultured Wharton’s Jelly-derived mesenchymal stem cells can achieve higher electroporation efficiency via two pulse sessions at a voltage of 150 V, pulse duration of 5.0 ms, and pulse interval time of 50 ms.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Umbilical Cord, Mesenchymal Stem Cells, Transfection, Tissue Engineering

CLC Number:

R394.2

Wang Ze, Li Chun-hua, Zhang Ning-kun, Gao Lian-ru, Chen Yu. Optimal conditions for transfecting Wharton’s Jelly-derived mesenchymal stem cells with electroporation method[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(17): 2717-2721.

Figures/Tables 1

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