Orth Patrick, Weimer Anja, Kaul Gunter, Kohn Dieter, Cucchiarini Magali, Madry Henning
Laboratory for Experimental Orthopaedics, Department of Orthopaedic Surgery, Saarland University Medical Center, Homburg, Germany.
Mol Biotechnol. 2008 Feb;38(2):137-44. doi: 10.1007/s12033-007-0071-8. Epub 2007 Oct 17.
The aim of the present study was to evaluate the efficacy of novel nonviral gene delivery systems in cells of musculoskeletal origin. Primary cultures of lapine skeletal muscle cells, lapine articular chondrocytes, human cells from fibrous dysplasia and cell lines established from human osteosarcoma (SAOS-2), chondrosarcoma (CS-1), murine skeletal myoblasts (L8) and fibroblasts (NIH 3T3) were transfected with the P. pyralis luc or the E. coli lacZ genes using Nanofectin 1 and 2, Superfect, JetPEI, GeneJammer, Effectene, TransPass D2, FuGENE 6, Lipofectamine 2000, Dreamfect, Metafectene, Escort III, and calcium phosphate. Maximal transfection efficiency in lapine skeletal muscle cells was of 60.8 +/- 21.2% using Dreamfect, 38.9 +/- 5.0% in articular chondrocytes using Gene Jammer, 5.2 +/- 8.0% in human cells from fibrous dysplasia using Lipofectamine 2000, 12.7 +/- 16.2% in SAOS-2 cells using FuGENE 6, 29.9 +/- 3.5% in CS-1 cells using Lipofectamine 2000, 70.7 +/- 8.6% in L8 cells using FuGENE 6, and 48.9 +/- 13.0% in NIH 3T3 cells using Metafectene. When the cells were transfected with a human IGF-I gene, significant amounts of the IGF-I protein were secreted. These results indicate that relatively high levels of transfection can be achieved using novel nonviral gene transfer methods.
本研究的目的是评估新型非病毒基因递送系统在肌肉骨骼来源细胞中的疗效。使用Nanofectin 1和2、Superfect、JetPEI、GeneJammer、Effectene、TransPass D2、FuGENE 6、Lipofectamine 2000、Dreamfect、Metafectene、Escort III和磷酸钙,将萤火虫荧光素酶基因或大肠杆菌β-半乳糖苷酶基因转染至兔骨骼肌细胞、兔关节软骨细胞、骨纤维发育不良的人细胞以及源自人骨肉瘤(SAOS-2)、软骨肉瘤(CS-1)、小鼠骨骼肌成肌细胞(L8)和成纤维细胞(NIH 3T3)的细胞系。使用Dreamfect时,兔骨骼肌细胞的最大转染效率为60.8±21.2%;使用Gene Jammer时,关节软骨细胞的最大转染效率为38.9±5.0%;使用Lipofectamine 2000时,骨纤维发育不良的人细胞的最大转染效率为5.2±8.0%;使用FuGENE 6时,SAOS-2细胞的最大转染效率为12.7±16.2%;使用Lipofectamine 2000时,CS-1细胞的最大转染效率为29.9±3.5%;使用FuGENE 6时,L8细胞的最大转染效率为70.7±8.6%;使用Metafectene时,NIH 3T3细胞的最大转染效率为48.9±13.0%。当用人类IGF-I基因转染细胞时,可分泌大量的IGF-I蛋白。这些结果表明,使用新型非病毒基因转移方法可实现相对较高水平的转染。