Regeneration of neurons and vessels using bone marrow cells and gene delivery

我们初步将年轻小鼠骨髓细胞静脉植入老年加速衰老小鼠（SAM）体内。水迷宫对大鼠的记忆功能没有明显改善。因此，静脉注射年轻骨髓对预防衰老和进一步加强治疗（如有用的基因诱导和靶向给药到大脑或骨髓）的作用不大。然后，我们通过单纯疱疹病毒1型（HSV-1）载体转染FGF-2基因间充质基质细胞（MSCs），研究了一种新的治疗策略对短暂性大脑中动脉闭塞（MCAO）后大鼠神经系统改善的影响。与其他组相比，接受fgf -2修饰MSCs的中风动物表现出显著的功能恢复。MCAO 14天后，仅fgf -2修饰的mscs处理组梗死体积显著减少。MCAO后3天和7天，FGF-2修饰的mscs -t脑组织中FGF-2的生成明显高于其他组。经酶联免疫吸附试验证实，FGF-2修饰的MSCs表达FGF-2蛋白。总之，这些数据表明，与单独移植MSCs相比，带有HSV-1载体的FGF-2基因修饰的MSCs有助于卒中后功能恢复。肝细胞生长因子（HGF）也是组织修复的有用基因之一。我们介绍了一种利用多突变HSV-1载体在大鼠瞬时MCAO模型中结合MSCs和离体HGF基因转移的新策略。超急性治疗时，仅mscs - hgf组与PBS组在第14天观察到梗死面积的显著差异，两种移植均在第14天观察到梗死面积的显著差异。在超急性移植后，我们在治疗后第1天检测到MSC-HGF组缺血脑中HGF蛋白的表达较其他组丰富，并维持至少2周。此外，我们确定HGF表达的增加来源于基因修饰的MSCs中转移的HGF基因。与其他组相比，MSC-HGF组缺血边界区（IBZ）凋亡阳性细胞比例显著降低，IBZ皮层剩余神经元比例显著升高。目前的研究表明，联合治疗比单独MSC细胞治疗更有效，并且可以延长治疗时间窗口，从超急性期到急性期。通过这些实验，改善效果主要是由于抗凋亡和神经元保护作用，没有证据表明神经元生成。今后还应研究骨髓内注射骨髓细胞等其他有效方法。少

We preliminarily intravenously implanted bone marrow cells from young mouse to old senescence accelerated mouse (SAM). We evaluate the memory function by water maze, however, no improvement in memory function was observed. Therefore, intravenous administration of young bone marrow has little effect on preventing aging and further enhanced treatment such as useful gene induction and targeting administration to the brain or bone marrow.Then we examined the effects of neurological improvement after transient middle cerebral artery occlusion (MCAO) in rats by a novel therapeutic strategy with FGF-2 gene-transferred mesenchymal stromal cells (MSCs) by the herpes simplex virus type 1 (HSV-1) vector. The stroke animals receiving FGF-2-modified MSCs demonstrated significant functional recovery compared with the other groups. Fourteen days after the MCAO, there was a significant reduction in infarction volume only in FGF-2-modified MSC-treated group. FGF-2 production in the FGF-2-modified MSC-t … More reated brain was significantly higher compared with the other groups at 3 and 7 days after MCAO. Administrated FGF-2-modified MSCs strongly expressed the FGF-2 protein, which was proven by ELISA. In conclusion these datum suggested that the FGF-2 gene-modified MSCs with the HSV-1 vector can contribute to remarkable functional recovery after stroke compared with MSCs transplantation alone.The hepatocyte growth factor (HGF) is also one of the useful gene for tissue restoration. we introduce a new strategy combining MSCs and ex vivo HGF gene transferring with a multimutated HSV-1 vector in a rat transient MCAO model. The significant difference of infarction areas on day was detected only between the MSC-HGF group and the PBS group with the superacute treatment, but was detected among each group on day 14 with both transplantations. After the superacute transplantation, we detected abundant expression of HGF protein in the ischemic brain of the MSC-HGF group compared with others on day 1 after treatment, and it was maintained for at least 2 weeks. Furthermore, we determined that the increased expression of HGF was derived from the transferred HGF gene in gene-modified MSCs. The percentage of apoptosis-positive cells in the ischemic boundary zone (IBZ) was significantly decreased, while that of remaining neurons in the cortex of the IBZ was significantly increased in the MSC-HGF group compared with others. The present study shows that combined therapy is more therapeutically efficient than MSC cell therapy alone, and it may extend the therapeutic time window from superacute to acute phase.Following these experiments, the improving effect is mainly due to anti-apoptotic and neuron-protective effects and there were no evidence for neuron-generation. Another effective methods such as intra-bone marrow injection of bone marrow cell should be examined in the future. Less