中国组织工程研究2018，Vol.22Issue(17)：2717-2721,5.DOI:10.3969/j.issn.2095-4344.0870
电穿孔法转染脐带华通胶间充质干细胞的最佳条件
Optimal conditions for transfecting Wharton's Jelly-derived mesenchymal stem cells with electroporation method
摘要
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.关键词
电穿孔/电转染/华通胶间充质干细胞/华通胶/低氧培养/干细胞/国家自然科学基金分类
医药卫生引用本文复制引用
王泽,李春花,张宁坤,高连如,陈宇..电穿孔法转染脐带华通胶间充质干细胞的最佳条件[J].中国组织工程研究,2018,22(17):2717-2721,5.基金项目
国家自然科学基金(81370238,81370901) the National Natural Science Foundation of China, No. 81370238, 81370901 （81370238,81370901）
