Mosahebi A, Woodward B, Wiberg M, Martin R, Terenghi G
Blond McIndoe Laboratories, University Department of Surgery, Royal Free and University College Medical School, London, England.
Glia. 2001 Apr 1;34(1):8-17. doi: 10.1002/glia.1035.
Transplantation of Schwann cells (SCs) is a promising treatment modality to improve neuronal regeneration. Identification of the transplanted cells is an important step when studying the development of this method. Genetic labeling is the most stable and reliable method of cell identification, but it is still unclear whether it has deleterious effect on SC characteristics. Our aim was to achieve a stable population of SCs transduced with the lacZ gene at a high frequency using a retroviral vector in vitro, and to follow the labeled SC in vitro to assess their viability and phenotypic marker expression. Furthermore, we transplanted lacZ-labeled SCs in a conduit to repair peripheral nerve to investigate their effect on nerve regeneration in vivo. Rat and human SCs were cultured and transduced with an MFG lacZ nls marker gene, achieving a transduction rate of 80% and 70%, respectively. Rat SCs were kept in culture for 27 weeks and examined every 4 weeks for expression of lacZ, viability, and phenotypic marker expression of GFAP, p75, MHC I and II. Throughout this period, transduced rat SCs remained viable and continued to proliferate. The proportion of cells expressing lacZ dropped only by 10% and the expression of phenotypic markers remained stable. Transduced human SCs were followed up for 4 weeks in culture. They proliferated and continued to express the lacZ gene and phenotypic marker expression of GFAP and p75 was preserved. Primary culture of transduced rat SCs were transplanted, syngeneically, in a conduit to bridge a 10 mm gap in sciatic nerve and the grafts were examined after 3 weeks for the presence and participation of labeled SCs and for axonal regeneration distance. Transplanted transduced rat SCs were clearly identified, taking part in the regeneration process and enhancing the axonal regeneration rate by 100% (at the optimal concentration) compared to conduits without SCs. Thus, retroviral introduction of lacZ gene has no deleterious effect on SCs in vitro and these SCs take part and enhance nerve regeneration in vivo.
雪旺细胞（SCs）移植是一种很有前景的改善神经元再生的治疗方式。在研究该方法的发展过程中，识别移植细胞是重要的一步。基因标记是最稳定、可靠的细胞识别方法，但尚不清楚其是否会对雪旺细胞特性产生有害影响。我们的目的是在体外使用逆转录病毒载体以高频转导携带lacZ基因的雪旺细胞群，并在体外追踪标记的雪旺细胞以评估其活力和表型标志物表达。此外，我们将用lacZ标记的雪旺细胞移植到导管中修复周围神经，以研究它们在体内对神经再生的影响。培养大鼠和人类雪旺细胞并用MFG lacZ nls标记基因进行转导，转导率分别达到80%和70%。将大鼠雪旺细胞培养27周，每4周检测一次lacZ表达、活力以及胶质纤维酸性蛋白（GFAP）、p75、主要组织相容性复合体I类和II类的表型标志物表达。在此期间，转导的大鼠雪旺细胞保持活力并继续增殖。表达lacZ的细胞比例仅下降了10%，表型标志物的表达保持稳定。转导的人类雪旺细胞在培养中随访4周。它们增殖并继续表达lacZ基因，GFAP和p75的表型标志物表达得以保留。将转导的大鼠雪旺细胞原代培养物同基因移植到导管中以桥接坐骨神经10毫米的间隙，3周后检查移植物中标记雪旺细胞的存在和参与情况以及轴突再生距离。移植的转导大鼠雪旺细胞清晰可辨，参与再生过程，与无雪旺细胞的导管相比，（在最佳浓度下）轴突再生率提高了100%。因此，逆转录病毒介导的lacZ基因导入在体外对雪旺细胞没有有害影响，并且这些雪旺细胞在体内参与并促进神经再生。
