Three-dimensional spheroids of adipose-derived mesenchymal stem cells are potent initiators of blood vessel formation in porous polyurethane scaffolds
Three-dimensional spheroids of adipose-derived mesenchymal stem cells are potent initiators of blood vessel formation in porous polyurethane scaffolds
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DOI：
10.1016/j.actbio.2013.02.013
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发表时间：
2013-06-01
影响因子：
9.7
通讯作者：
Menger, M. D.
中科院分区：
文献类型：
--
作者：
Laschke, M. W.;Schank, T. E.;Menger, M. D.
脂肪间充质干细胞（adMSCs）具有较高的血管生成活性。因此，将它们掺入组织构建体中代表了组织工程中有前途的血管化策略。在本研究中，我们分析了这种方法的功效是否可以通过将adMSCs作为三维球体接种到多孔支架上来提高。从C57 BL/6-TgN（ACTB-EGFP）1 Osb/J小鼠中分离表达CD 13、CD 73、CD 90和CD 117的绿色荧光蛋白（GFP）阳性adMSC，用于使用液体覆盖技术产生球状体。将这些球状体或相当数量的单个adMSC接种到多孔聚氨酯支架中，并植入C57 BL/6野生型小鼠的背侧皮褶室中。通过活体荧光显微镜和免疫组织化学分析植入物的血管化，并与非接种支架进行比较。adMSC球状体表现出类似于270 μ m的均匀直径，并且可以通过动态接种容易地并入支架中。植入后，它们诱导了强烈的血管生成宿主组织反应，导致支架血管化改善，与未接种的支架和接种有单个adMSC的支架相比，功能性微血管密度显著更高。免疫组织化学分析显示，一个高分数的类似于40%的所有微血管的中心内的球状体接种的支架从GFP阳性的adMSCs开发。这些血管与宿主的向内生长的GFP阴性血管融合。这表明adMSC球状体作为单独的血管化单元，促进在植入的组织构建体内的不同位置处新的微血管网络的同时发育。因此，adMSC球状体可用于增加基于MSC的血管化策略在未来组织工程应用中的功效。(C)2013 Acta Materialia Inc.由爱思唯尔有限公司出版。保留所有权利。
Adipose-derived mesenchymal stem cells (adMSCs) exhibit a high angiogenic activity. Accordingly, their incorporation into tissue constructs represents a promising vascularization strategy in tissue engineering. In the present study, we analyzed whether the efficacy of this approach can be improved by seeding adMSCs as three-dimensional spheroids onto porous scaffolds. Green fluorescent protein (GFP)-positive adMSCs expressing CD13, CD73, CD90 and CD117 were isolated from C57BL/6-TgN(ACTB-EGFP)1Osb/J mice for the generation of spheroids using the liquid overlay technique. Porous polyurethane scaffolds were seeded with these spheroids or a comparable number of individual adMSCs and implanted into the dorsal skinfold chamber of C57BL/6 wild-type mice. The vascularization of the implants was analyzed and compared to non-seeded scaffolds by means of intravital fluorescence microscopy and immunohistochemistry. The adMSC spheroids exhibited a homogeneous diameter of similar to 270 mu m and could easily be incorporated into the scaffolds by dynamic seeding. After implantation, they induced a strong angiogenic host tissue response, resulting in an improved scaffold vascularization with a significantly higher functional microvessel density when compared to non-seeded scaffolds and scaffolds seeded with individual adMSCs. Immunohistochemical analyses revealed that a high fraction of similar to 40% of all microvessels within the center of spheroid-seeded scaffolds developed from GFP-positive adMSCs. These vessels inosculated with ingrowing GFP-negative vessels of the host. This indicates that adMSC spheroids serve as individual vascularization units, promoting the simultaneous development of new microvascular networks at different locations inside implanted tissue constructs. Thus, adMSC spheroids may be used to increase the efficacy of MSC-based vascularization strategies in future tissue engineering applications. (C) 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.