Characterisation of the interaction between circulating and in vitro cultivated endothelial progenitor cells and the endothelial barrier
Characterisation of the interaction between circulating and in vitro cultivated endothelial progenitor cells and the endothelial barrier
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DOI：
10.1016/j.ejcb.2007.08.002
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发表时间：
2008-02-01
影响因子：
6.6
通讯作者：
Bloch, Wilhelm
中科院分区：
文献类型：
--
作者：
Funcke, Fabienne;Hoyer, Heike;Bloch, Wilhelm
体外培养内皮祖细胞(CEPC)用于心脏再生的冠脉内细胞治疗。本研究的目的是研究cEPC和循环中的单个核细胞(MNC)是否能够通过内皮屏障进入心脏组织。从健康男性志愿者(n=13，25+/-6岁)外周血中分离单个核细胞(MNC)和内皮祖细胞(EPC)，并用荧光标记进行染色。这些细胞通过含有不同表型的内皮层的器官(大鼠主动脉、人脐静脉、离体鼠心脏)在体外灌流。对内皮和基底膜进行免疫荧光染色，冰冻切片用激光共聚焦扫描显微镜进行分析。经大鼠主动脉灌流后，MNC在内皮层和内皮细胞下的基底层有黏附/整合。然而，没有发现MNC在内皮屏障上的迁移。即使细胞数量增加(从0.5百万个/小时增加到1000万个/小时)、延长灌流时间(1.5-4h)和培养24小时，这一点仍然成立。在含完整内皮细胞的朗宁多夫灌流小鼠心脏中，MNC(1×10(7))和cEPC(1×10(6))在0.5和2小时后均未观察到迁移，说明MNC和cEPC不具有迁移内皮屏障的能力。因此，在干细胞治疗的背景下，这些细胞可能是内皮再生者，而不是心肌细胞的替代品。(C)2007年爱思唯尔股份有限公司。版权所有。
In vitro cultured endothelial progenitor cells (cEPC) are used for intracoronary cell therapy in cardiac regeneration. The aim of this study was to investigate whether cEPC and circulating mononuclear cells (MNC), which include a small number of in vivo circulating EPC, are able to transmigrate through the endothelial barrier into the cardiac tissue. MNC and EPC were isolated from the peripheral blood from healthy male volunteers (n = 13, 25 +/- 6 years) and stained with a fluorescent marker. The cells were perfused in vitro through organs with endothelial layers of different phenotypes (rat aorta, human umbilical vein, isolated mouse heart). The endothelium and the basal lamina were then stained by immunofluorescence and the cryo-sections analysed using a confocal laser scanning microscope. After perfusion through the rat aorta, an adhesion/integration of MNC was observed at the endothelial layer and the basal lamina beneath endothelial cells. However, no migration of MNC over the endothelial barrier was found. This remained true even when the cell numbers were increased (from 0.5 to 10 million cells/h), when the time of perfusion was prolonged (1.5-4h) and when the aorta was cultivated for 24h. In the Langendorff-perfused mouse heart with intact endothelium, no migration of MNC (1 x 10(7)) or cEPC (1 x 10(6)) was observed after 0.5 and 2 h. In conclusion, MNC and cEPC do not possess any capacity to transmigrate the endothelial barrier. In the context of stem cell therapy, these cells may therefore serve as endothelial regenerators but not as cardiomyocyte substitutes. (c) 2007 Elsevier GmbH. All rights reserved.