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

间充质干细胞（MSCs）在体内和体外均可分化为成骨细胞、神经细胞、骨骼肌细胞和血管内皮细胞。有报道称骨髓间充质干细胞植入可诱导缺血肢体血管生成。我们评估了骨髓间充质干细胞植入对肢体缺血家兔肢体血流（LBF）对乙酰胆碱（ACh）（一种内皮依赖性血管扩张剂）和硝普钠（SNP）（一种内皮依赖性血管扩张剂）反应的影响，将培养的骨髓间充质干细胞植入或注射生理盐水作为对照组。用电磁流量计测量LBF。每条缺血肢体共植入10^6个mscs。缺血肌肉组织学切片显示，骨髓间充质干细胞植入组毛细血管指数明显高于对照组。骨髓间充质干细胞植入组激光多普勒血流灌注指数明显高于对照组。MSC组LBF对乙酰胆碱的反应大于对照组。给予N^ g -硝基- l -精氨酸后，骨髓间充质干细胞植入组和对照组LBF对乙酰胆碱的反应相似。SNP在两组的血管舒张作用相似。在兔缺血模型中，骨髓间充质干细胞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。BM-MNC植入术对于严重肢体缺血的患者是有用的。然而，在BM-MNC抽吸过程中，患者接受全身麻醉。本研究建立了自体间充质干细胞培养方法。我们计划将培养的骨髓间充质干细胞储存，并在肢体缺血患者中进行自体骨髓间充质干细胞植入。

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.