Endothelial cells derived from human iPSCS increase capillary density and improve perfusion in a mouse model of peripheral arterial disease.
Endothelial cells derived from human iPSCS increase capillary density and improve perfusion in a mouse model of peripheral arterial disease.
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DOI：
10.1161/atvbaha.111.230938
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发表时间：
2011-11
期刊：
影响因子：
--
通讯作者：
Cooke JP
中科院分区：
文献类型：
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作者：
Rufaihah AJ;Huang NF;Jamé S;Lee JC;Nguyen HN;Byers B;De A;Okogbaa J;Rollins M;Reijo-Pera R;Gambhir SS;Cooke JP
已经使用成体或胚胎干细胞研究了用于血管生成和血管再生的干细胞疗法。在本研究中，我们研究了来自人诱导多能干细胞（hiPSC）的内皮细胞（EC）促进外周动脉疾病小鼠模型中缺血组织灌注的潜力。通过在补充有BMP-4和VEGF的分化培养基中培养hiPSC 14天来启动内皮分化。hiPSC-EC通过在基质胶中形成毛细血管样结构并掺入乙酰化LDL而表现出内皮特征。EC标志物如KDR、CD 31、CD 144和eNOS染色阳性。在体外暴露的hiPSC-EC缺氧导致各种血管生成相关的细胞因子和生长因子的表达增加。用由泛素启动子编码的双融合构建体、用于生物发光成像（BLI）的萤火虫荧光素酶和用于荧光检测的绿色荧光蛋白（GFP）（pUb-Fluc-GFP）稳定转导hiPSC-EC。将hiPSC-EC（5×105）在股动脉结扎后第0天和第7天通过肌内注射递送到SCID小鼠的缺血后肢中（n=8）。BLI显示hiPSC-EC在缺血肢体中存活至少2周。此外，激光多普勒成像显示，与盐水处理组相比，hiPSC-EC处理组的血液灌注率增加（0.58±0.12 vs 0.44±0.04; P=0.005）。接受hiPSC-EC注射的小鼠的缺血肢体中的毛细血管总数大于盐水处理组中的毛细血管总数（1284 ±155 vs. 797±206毛细血管/mm 2）（P<0.002）。这项研究是基于使用来自hiPSC的EC开发外周动脉疾病再生策略的第一步。
Stem cell therapy for angiogenesis and vascular regeneration has been investigated using adult or embryonic stem cells. In the present study, we investigated the potential of endothelial cells (ECs) derived from human induced pluripotent stem cells (hiPSCs) to promote the perfusion of ischemic tissue in a murine model of peripheral arterial disease. Endothelial differentiation was initiated by culturing hiPSCs for 14 days in differentiation media supplemented with BMP-4 and VEGF. The hiPSC-ECs exhibited endothelial characteristics by forming capillary-like structures in matrigel and incorporating acetylated-LDL. They stained positively for EC markers such as KDR, CD31, CD144 and eNOS. In vitro exposure of hiPSC-ECs to hypoxia resulted in increased expression of various angiogenic related cytokines and growth factors. hiPSC-ECs were stably transduced with a double fusion construct encoded by the ubiquitin promoter, firefly luciferase for bioluminescence imaging (BLI) and green fluorescence protein (GFP) for fluorescent detection (pUb-Fluc-GFP). The hiPSC-ECs (5×105) were delivered by intramuscular injection into the ischemic hindlimb of SCID mice at day 0 and again on day 7 after femoral artery ligation (n=8). BLI showed that hiPSC-ECs survived in the ischemic limb for at least 2 weeks. In addition, laser Doppler imaging showed that the ratio of blood perfusion was increased by hiPSC-EC treatment by comparison to the saline-treated group (0.58±0.12 vs 0.44±0.04; P=0.005). The total number of capillaries in the ischemic limb of mice receiving hiPSC-EC injections was greater than those in the saline-treated group (1284 ±155 vs. 797±206 capillaries/mm2) (P<0.002). This study is a first step toward development of a regenerative strategy for peripheral arterial disease based on the use of ECs derived from hiPSCs.