Microvessel-like structures from outgrowth endothelial cells from human peripheral blood in 2-dimensional and 3-dimensional co-cultures with osteoblastic lineage cells.

Sabine Fuchs, Alexander Hofmann, C James Kirkpatrick
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引用次数: 166

Abstract

Tissue regeneration involves complex processes in the interaction between different cell types that control the process of neo-vascularization. In bone, osteoblasts and bone marrow stem cells provide cue elements for the proliferation of endothelial cells, differentiation of endothelial precursors, and the maturation of a vascular network. In this study, we investigated outgrowth endothelial cells (OECs), a potential source of autologous endothelial cells derived from human peripheral blood, in direct 2-dimensional (2-D) and 3-D co-culture systems with cells relevant for the regeneration of bone tissue, such as osteoblasts. In the co-cultures, OECs were evaluated in terms of their stability as an endothelial population at the single cell level using flow cytometry and their ability to establish a pre-vascular network at the light-microscopical and ultra-structural level. In co-cultures with the osteoblast cell line MG63 and with human primary osteoblasts (pOBs), OECs, in contrast to human umbilical vein endothelial cells, formed highly organized microvessel-like structures. These microvessel-like structures included the formation of a vascular lumen with tight junctional complexes at intercellular contacts of endothelial cells. In the co-culture, the formation of this vascular network was achieved in the standard growth medium for OECs. Furthermore, using a rotating culture vessel system, 3-D co-cultures consisting of OECs and pOBs were generated. Based on these observations, we conclude that OECs could provide a valuable source of autologous endothelial cells for the generation of complex tissue-engineered tissues.

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人外周血内皮细胞与成骨细胞系在二维和三维共培养中的微血管样结构。
组织再生涉及控制新血管形成过程的不同细胞类型之间相互作用的复杂过程。在骨中,成骨细胞和骨髓干细胞为内皮细胞的增殖、内皮前体的分化和血管网络的成熟提供线索。在这项研究中,我们研究了外生内皮细胞(OECs),一种来源于人外周血的自体内皮细胞的潜在来源,在直接的二维(2d)和三维共培养系统中与骨组织再生相关的细胞(如成骨细胞)共培养。在共培养中,使用流式细胞术评估oec在单细胞水平上作为内皮细胞群的稳定性,以及它们在光学显微镜和超结构水平上建立血管前网络的能力。在与成骨细胞系MG63和人原代成骨细胞(pob)共培养中,与人脐静脉内皮细胞相比,oec形成了高度组织化的微血管样结构。这些微血管样结构包括在内皮细胞的细胞间接触处形成具有紧密连接复合物的血管腔。在共培养中,这种维管网络的形成是在oec的标准生长培养基中实现的。此外,使用旋转培养容器系统,生成由oec和pob组成的三维共培养物。基于这些观察结果,我们得出结论,oec可以为复杂组织工程组织的生成提供有价值的自体内皮细胞来源。
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Tissue engineering
Tissue engineering CELL & TISSUE ENGINEERING-BIOTECHNOLOGY & APPLIED MICROBIOLOGY
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