Early endothelial progenitor cells as a source of myeloid cells to improve the pre-vascularisation of bone constructs.
Early endothelial progenitor cells as a source of myeloid cells to improve the pre-vascularisation of bone constructs.
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DOI：
10.22203/ecm.v027a06
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发表时间：
2014-01
影响因子：
3.1
通讯作者：
Y. Shi;G. Kramer;A. Schröder;C. Kirkpatrick;A. Seekamp;H. Schmidt;S. Fuchs
中科院分区：
文献类型：
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作者：
Y. Shi;G. Kramer;A. Schröder;C. Kirkpatrick;A. Seekamp;H. Schmidt;S. Fuchs
根据目前的知识，血源性内皮祖细胞（EPC）可能具有血管生成前骨髓细胞的作用，在血管生成和血管重组的调节中起着重要作用。在这种情况下，我们评估了内源性髓细胞在血源性生长内皮细胞（OEC）和成骨细胞共培养中的作用。此外，我们在由间充质干细胞（MSC）和间充质干细胞（OEC）组成的体外模型中研究了EPC作为髓系细胞形成血管结构的潜在来源的作用。为此，我们将EPCs添加到MSC和oec共培养中。通过共聚焦激光显微镜（CLSM）分析血管结构和髓系细胞的共定位，用于内皮和髓系标记和定量图像分析。通过实时荧光定量PCR、酶联免疫吸附及细胞培养上清和裂解物的蛋白阵列分析骨髓细胞的分子效应。在OEC和成骨细胞共培养中，内源性髓细胞与血管生成结构显著共定位。在OEC和MSC共培养过程中主动添加EPC，在共培养过程的早期阶段，统计上证实了血管前结构形成的增加。此外，我们观察到内皮标记物的增加，表明EPC或髓样细胞对内皮细胞生长有有益作用。此外，real - time PCR显示，在EPC和MSC共培养中，CD68、CD11b和CD163的表达水平较高，表明EPC至少在一定程度上具有巨噬细胞样细胞的作用。
According to present knowledge, blood derived endothelial progenitor cells (EPC) might act as proangiogenic myeloid cells, which play a fundamental role in the regulation of angiogenesis and blood vessel reorganisation. In this context, we have evaluated the contribution of endogenous myeloid cells in co-cultures of blood derived outgrowth endothelial cells (OEC) and osteogenic cells. In addition, we investigated the role of EPC as a potential source of myeloid cells in the formation of vascular structures in an in vitro model consisting of mesenchymal stem cells (MSC) and OEC. For this purpose, we added EPCs to co-cultures of MSC and OECs. Vascular structures and the co-localisation of myeloid cells were analysed by confocal laser microscopy (CLSM) for endothelial and myeloid markers and quantitative image analysis. The molecular effects of myeloid cells were evaluated by quantitative real time PCR, ELISA and protein arrays from cell culture supernatants and lysates. Endogenous myeloid cells were significantly co-localised with angiogenic structures in co-cultures of OEC and osteogenic cells. The active addition of EPC to co-cultures of OEC and MSC resulted in a statistically approved increase in the formation of prevascular structures at early stages of the co-culture process. In addition, we observed an increase of endothelial markers, indicating beneficial effects of EPC or myeloid cells on endothelial cell growth. Furthermore, real time PCR indicated high expression levels of CD68, CD11b and CD163 in co-cultures of EPC and MSC indicating that EPC act at least partly as macrophage like-cells.