Substrate-dependent Wnt signaling in MSC differentiation within biomaterial-derived 3D spheroids
Substrate-dependent Wnt signaling in MSC differentiation within biomaterial-derived 3D spheroids
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DOI：
10.1016/j.biomaterials.2013.03.031
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发表时间：
2013-07-01
期刊：
影响因子：
14
通讯作者：
Huang, Guo-shiang
中科院分区：
文献类型：
--
作者：
Hsu, Shan-hui;Huang, Guo-shiang
开发了一种独特的基于生物材料的系统来生成动态三维(3D)多细胞球体的间充质干细胞(MSCs)。将MSCs培养在不同密度的壳聚糖透明膜或透明质酸(HA)透明膜上。MSCs在基质依赖性上调黏附分子N-钙粘附素的作用下，迁移旺盛，自组装成高流动性的三维球体。MSC球体显示Wnt基因/蛋白表达增加，细胞命运依赖于底物。在MSC球体中观察到分化能力与Wnt信号和与ERK1/2或Smad2/3等其他信号通路的串扰相关，而在常规2D培养细胞中则未观察到相关。WNT3a介导的规范Wnt信号在壳聚糖衍生的MSC球体中更为活跃，易于成骨。Wnt5A介导的非规范Wnt信号对于在HA接枝的壳聚糖上形成的MSC球体更活跃，容易形成软骨。特别是，WNT5a介导的非正则Wnt信号与WNT3a介导的正则Wnt信号在决定细胞命运中的相对重要性受HA在壳聚糖上的接枝密度控制。经规范的Wnt相关信号分子抑制剂处理后，MSC球体在壳聚糖上的成骨受到抑制。本研究表明，骨髓间充质干细胞的Wnt信号在3D环境中是不同的，并且是底物依赖的。这个方便的3D平台可以用来研究Wnt信号在不同细胞外环境下控制MSC命运的作用，并有可能应用于再生医学、正常发育和癌症中干细胞行为的研究。(C)2013爱思唯尔有限公司。保留所有权利。
A unique biomaterial-based system was developed to generate dynamic three-dimensional (3D) multicellular spheroids of mesenchymal stem cells (MSCs). MSCs were cultured on transparent membranes made of chitosan or those further grafted with hyaluronan (HA) in different densities. MSCs vigorously migrated and were self-assembled into highly mobile 3D spheroids with substrate-dependent upregulation of adhesion molecule N-cadherin. MSC spheroids showed increased expression of Wnt genes/proteins and substrate-dependent cell fate. The correlation of differentiation capacities with Wnt signaling and crosstalk with other pathways such as ERK1/2 or Smad2/3 were observed for MSC spheroids but not for the conventional 2D cultured cells. Wnt3a-mediated canonical Wnt signaling was more active for MSC spheroids derived on chitosan, which were prone to osteogenesis. Wnt5a-mediated non-canonical Wnt signaling was more active for MSC spheroids derived on HA-grafted chitosan, which were prone to chondrogenesis. In particular, the relative importance of Wnt5a-mediated non-canonical vs. Wnt3a-mediated canonical Wnt signals in determining the cell fate was controlled by the grafting density of HA on chitosan. Treatment with the inhibitor of canonical Wnt-associated signaling molecules suppressed the osteogenesis of MSC spheroids on chitosan. This study demonstrates that Wnt signaling of MSCs is distinct in 3D environment and is substrate-dependent. The convenient 3D platform may be used to examine the role of Wnt signaling in controlling MSC fate under different extracellular environments, and potentially applied to study stem cell behavior in regenerative medicine, normal development, and cancer. (C) 2013 Elsevier Ltd. All rights reserved.