Nakashima S, Matsuyama Y, Nitta A, Sakai Y, Ishiguro N
Department of Orthopaedic Surgery, Nagoya University School of Medicine, 65 Tsuruma-cho, Showa-ku, Nagoya 466-8550, Japan.
Transplant Proc. 2005 Jun;37(5):2290-2. doi: 10.1016/j.transproceed.2005.03.047.
Human marrow stromal cells (hMSCs) are an attractive source for autologous cell and gene therapies. In this study, we developed a highly efficient transfection method for hMSCs. Although they tend to show efficient gene delivery, nonviral vectors offer several advantages over viral vectors for gene therapies. They are inexpensive to produce and suitable to adopt particularly with respect to little or no specific immune responses; they are simple to use; they entail easier large-scale production; and they have a high degree quality control. hMSCs and rat marrow stromal cells were transfected with the plasmid pEGFP-N1 that encoded a green fluorescent protein component by using two nonviral methods: nucleofection and electroporation. Nucleofection provided a much better rate of transfer than electroporation particularly in hMSCs.
人骨髓基质细胞(hMSCs)是自体细胞和基因治疗的一个有吸引力的来源。在本研究中,我们开发了一种用于hMSCs的高效转染方法。尽管非病毒载体往往显示出高效的基因递送,但与病毒载体相比,非病毒载体在基因治疗方面具有几个优势。它们生产成本低廉,尤其适合于几乎没有或没有特异性免疫反应的情况;使用简单;易于大规模生产;并且具有高度的质量控制。通过使用两种非病毒方法:核转染和电穿孔,用编码绿色荧光蛋白成分的质粒pEGFP-N1转染hMSCs和大鼠骨髓基质细胞。核转染比电穿孔提供了更好的转染率,尤其是在hMSCs中。