Hoffmann J, Glassford A J, Doyle T C, Robbins R C, Schrepfer S, Pelletier M P
Cardiothoracic Surgery, Stanford University School of Medicine, Stanford, CA 94305-5407, USA.
Thorac Cardiovasc Surg. 2010 Apr;58(3):136-42. doi: 10.1055/s-0029-1240758.
Bone marrow-derived mesenchymal stem cells (MSCs) are multipotent and secrete angiogenic factors, which could help patients with occlusive arterial diseases. We hypothesize that MSCs, in comparison to fibroblasts, survive better under hypoxic conditions in vitro and in vivo. MSCs and fibroblasts from L2G mice expressing firefly luciferase and GFP were cultured in normoxic and hypoxic conditions for 24 hours. In vitro cell viability was tested by detecting apoptosis and necrosis. MSCs released higher amounts of VEGF (281.1 +/- 62.6 pg/ml) under hypoxic conditions compared to normoxia (154.9 +/- 52.3 pg/ml, p = NS), but were less tolerant to hypoxia (45 +/- 7.9%) than fibroblasts (28.1 +/- 3.6%, p = NS). A hindlimb ischemia model was created by ligating the femoral artery of 18 FVB mice. After one week, 1 x 106 cells (MSCs, fibroblasts or saline) were injected into the limb muscles of each animal (n = 6 per group). Bioluminescence measurement to assess the viability of luciferase positive cells showed significant proliferation of MSCs on day four compared to fibroblasts (p = 0.001). Three weeks after cell delivery, the capillary to muscle fiber ratio of ischemic areas was analyzed. In the MSC group, vessel density was significantly higher than in the fibroblast or control group (0.5 +/- 0.08 and 0.3 +/- 0.03). Under hypoxia, MSCs produced more VEGF compared to normal conditions and MSC transplantation into murine ischemic limbs led to an increase in vessel density, although MSC survival was limited. This study suggests that MSC transplantation may be an effective and clinically relevant tool in the therapy of occlusive arterial diseases.
骨髓间充质干细胞（MSCs）具有多能性且能分泌血管生成因子，这有助于治疗闭塞性动脉疾病患者。我们假设，与成纤维细胞相比，MSCs在体外和体内的低氧条件下存活得更好。将表达萤火虫荧光素酶和绿色荧光蛋白的L2G小鼠的MSCs和成纤维细胞在常氧和低氧条件下培养24小时。通过检测细胞凋亡和坏死来测试体外细胞活力。与常氧条件（154.9±52.3 pg/ml，p=无统计学意义）相比，MSCs在低氧条件下释放的血管内皮生长因子（VEGF）量更高（281.1±62.6 pg/ml），但与成纤维细胞相比，其对低氧的耐受性更低（45±7.9%）（28.1±3.6%，p=无统计学意义）。通过结扎18只FVB小鼠的股动脉建立后肢缺血模型。一周后，将1×10⁶个细胞（MSCs、成纤维细胞或生理盐水）注射到每只动物的肢体肌肉中（每组n=6）。通过生物发光测量来评估荧光素酶阳性细胞的活力，结果显示与成纤维细胞相比，MSCs在第4天有显著增殖（p=0.001）。细胞注射三周后，分析缺血区域的毛细血管与肌纤维比例。在MSCs组中，血管密度显著高于成纤维细胞组或对照组（0.5±0.08和0.3±0.03）。在低氧条件下，与正常条件相比，MSCs产生更多的VEGF，并且将MSCs移植到小鼠缺血肢体中导致血管密度增加，尽管MSCs的存活有限。这项研究表明，MSCs移植可能是治疗闭塞性动脉疾病的一种有效且与临床相关的工具。
