Therapeutic Angiogenesis with Autologous Mesenchymal Stem Cell in Patients with Peripheral Artery Diseases: Mesenchymal Stem Cell Storing System

骨髓间充质干细胞（Mesenchymal stem cells，MSCs）在体内外可分化为成骨细胞、神经细胞、骨骼肌细胞和血管内皮细胞。已有报道称，MSC移植可诱导缺血肢体血管生成活跃。我们评估了MSC植入对肢体血流（LBF）对乙酰胆碱（ACh）（一种内皮依赖性血管扩张剂）和硝普钠（SNP）（一种内皮非依赖性血管扩张剂）的反应的影响。使用电磁流量计测量LBF。每侧缺血肢体共植入10^6MSCs。缺血肌肉组织切片显示，MSC植入组的毛细血管指数大于对照组。激光多普勒血流灌注指数在MSC植入组较对照组显著增加。MSC组LBF对ACh的反应大于对照组。注射N^G-硝基-L-精氨酸后，MSC植入组和对照组对ACh的LBF反应相似。SNP的扩血管作用在两组中相似。在兔缺血模型中，MSC植入通过增加一氧化氮的产生诱导血管生成并增强内皮依赖性血管舒张。显然，BM-MNC植入对严重肢体缺血患者是有用的。然而，患者在BM-MNC抽吸期间接受全身麻醉。本研究建立了自体骨髓间充质干细胞的培养方法。我们正在计划储存培养的MSC，并在肢体缺血患者中进行自体MSC植入。

Mesenchymal stem cells (MSCs) can differentiate into osteoblasts, nerve cells, skeletal muscle cells, and vascular endothelial cells in vivo and in vitro. It has been reported that MSC implantation induces active angiogenesis in ischemic limbs. We evaluated the effect of MSC implantation on limb blood flow (LBF) responses to acetylcholine (ACh), an endothelium-dependent vasodilator, and sodium nitroprusside (SNP), an endothelium-independent vasodilator, in rabbits with limb ischemia in which cultured MSC were implanted or saline was injected as a control group. LBF was measured using an electromagnetic flowmeter. A total of 10^6MSCs were implanted into each ischemic limb. Histological sections of ischemic muscle showed that capillary index was greater in the MSC implantation group than in the control group. Laser Doppler blood perfusion index was significantly increased in the MSC implantation group compared with that in the control group. LBF response to ACh was greater in the MSC group than in the control group. After administration of N^G-nitro-L-arginine, LBF response to ACh was similar in the MSC implantation group and control group. Vasodilatory effects of SNP in the two groups were similar. MSC implantation induces angiogenesis and augments endothelium-dependent vasodilation in a rabbit ischemic model through an increase in nitric oxide production.It is clear that BM-MNC implantation is useful for patients with severe limb ischemia. However, patients were subjected by general anesthesia during BM-MNC aspiration. In the present study, we established the autologous MSC culture method. We are making a plan to store cultured MSC and to perform the autologous MSC implantation in patients with limb ischemia.