Osteogenic potential of adipose-derived macrospheroids cocultured with CD11b+ monocytes
Osteogenic potential of adipose-derived macrospheroids cocultured with CD11b+ monocytes
复制标题
脂肪来源的大球体与 CD11b 单核细胞共培养的成骨潜力
DOI：
10.11607/jomi.4957
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发表时间：
2017
影响因子：
2
通讯作者：
*Shinji Kuroda
中科院分区：
文献类型：
--
作者：
Hidemi Nakata;Maiko Yamamoto;Emi Okada;Tomoko Nagayama;Shohei Kasugai;*Shinji Kuroda
目的：在潜在的基于细胞的治疗中，脂肪源性干细胞（ASCs）被认为是一种有前途的组织再生干细胞来源。尽管最近的许多临床试验研究了脂肪组织或ASC在移植中的应用，但尚未对生长中的大球状体（macrospheroid）的微观结构或ASC球体与单核细胞/巨噬细胞谱系的三维共培养进行分析。本研究的目的是分析小鼠源性ASC大球体的微观结构，以及这些大球体在三维环境下和与单核细胞共培养诱导钙化后的生长和成骨潜力。材料与方法：采用苏木精染色、伊红染色和原位杂交技术对小鼠源性ASC大球体的组织结构和多能性标记基因的表达进行分析。透射电镜观察ASC巨球显微结构，并分析细胞在巨球中的增殖情况。此外，在二维和三维环境中以及与单核细胞共培养诱导钙化后，评估了这些大球体的生长和成骨潜力。结果：Oct3/4、Nanog和Sox2在球体深部也有表达，表面表达量较高。球状中心内可见细胞增殖。球体微观结构的观察表明，在球体结构中产生了细胞外基质。将球体移植到羟基磷灰石椎间盘上，形成三维细胞生长，填充椎间盘。球体与单核细胞共培养可形成许多破骨细胞样的多核细胞，共培养3周后观察到钙化。结论：ASC球体具有较高的三维动态生长和成骨分化能力。此外，ASC球状体促进单核细胞向破骨细胞样细胞分化，这可能增强ASC球状体的成骨潜能。
Purpose: Among potential cell-based therapies, adipose-derived stem cells (ASCs) have been proposed as a promising source of stem cells for tissue regeneration. Although many recent clinical trials have investigated the use of adipose tissue or ASCs in transplantation, analysis of the microstructures of outgrowing macrosized spheroids (macrospheroids) or three-dimensional coculture of ASC spheroids and monocyte/macrophage lineages has not been performed. The aim of this study was to analyze the microstructures of murine-derived ASC macrospheroids and the growth and osteogenic potential of these macrospheroids in a three-dimensional environment and after calcification induction by coculture with monocytes. Materials and Methods: The histologic structures of murine-derived ASC macrospheroids and the expression of marker genes for multipotency within these macrospheroids were analyzed by hematoxylin and eosin staining and in situ hybridization. ASC macrospheroid microstructures were observed by transmission electron microscopy, and cell proliferation in the spheroids was analyzed. Additionally, the growth and osteogenic potential of these macrospheroids were assessed in two-dimensional and three-dimensional environments and after calcification induction by coculture with monocytes. Results: The expression of Oct3/4, Nanog, and Sox2 was detected even in the deep zone of spheroids, although higher expression was observed at the surface. Cell proliferation was detected within the spheroid centers. Observation of spheroid microstructure revealed extracellular matrix production within the spheroid architecture. Transplantation of a spheroid on the hydroxyapatite disc resulted in three-dimensional cell growth, filling the disc. Coculture of the spheroids with monocytes led to the formation of many osteoclast-like, multinucleated cells, and calcification was observed after 3 weeks of coculture. Conclusion: ASC spheroids exhibited high capacity for dynamic three-dimensional growth and osteogenic differentiation. Furthermore, ASC spheroids promoted monocyte differentiation into osteoclast-like cells, which may enhance the osteogenic potential of ASC spheroids.