Ruitenberg M J, Plant G W, Christensen C L, Blits B, Niclou S P, Harvey A R, Boer G J, Verhaagen J
Graduate School for Neurosciences Amsterdam, Netherlands Institute for Brain Research, Amsterdam, The Netherlands.
Gene Ther. 2002 Jan;9(2):135-46. doi: 10.1038/sj.gt.3301626.
Implantation of olfactory ensheathing glia (OEG) is a promising strategy to augment long-distance regeneration in the injured spinal cord. In this study, implantation of OEG following unilateral hemisection of the dorsal cervical spinal cord was combined with ex vivo gene transfer techniques. We report, to our knowledge for the first time, that purified cultures of primary OEG are capable of expressing a foreign gene following adenoviral (AdV) and lentiviral (LV) vector-mediated gene transfer. OEG implants subjected to AdV vector-mediated gene transfer expressed high levels of transgenic protein in both intact and lesioned spinal cord at 7 days after implantation. However, the levels of transgene expression gradually declined between 7 and 30 days after implantation in lesioned spinal cord. Infection with LV vectors resulted in stable transduction of primary OEG cultures and transgene expression persisted for at least 4 months after implantation. Genetic engineering of OEG opens the possibility of expressing additional neurotrophic genes and create optimal 'bridging' substrates to support spinal axon regeneration. Furthermore, stable transduction of OEG allows us to reliably study the behaviour of implanted cells and to obtain better understanding of their regeneration supporting properties.
植入嗅鞘胶质细胞（OEG）是增强脊髓损伤后长距离再生的一种有前景的策略。在本研究中，将颈段脊髓背侧半横断后植入OEG与体外基因转移技术相结合。据我们所知，我们首次报道，原代OEG的纯化培养物在腺病毒（AdV）和慢病毒（LV）载体介导的基因转移后能够表达外源基因。接受AdV载体介导基因转移的OEG植入物在植入后7天在完整和损伤的脊髓中均表达高水平的转基因蛋白。然而，在损伤脊髓中，植入后7至30天转基因表达水平逐渐下降。用LV载体感染导致原代OEG培养物的稳定转导，并且转基因表达在植入后持续至少4个月。OEG的基因工程开启了表达额外神经营养基因并创建最佳“桥接”底物以支持脊髓轴突再生的可能性。此外，OEG的稳定转导使我们能够可靠地研究植入细胞的行为，并更好地了解它们的再生支持特性。
