A quantitative approach for determining the role of geometrical constraints when shaping mesenchymal condensations.
A quantitative approach for determining the role of geometrical constraints when shaping mesenchymal condensations.
复制标题
一种确定间充质凝结成形时几何约束作用的定量方法。
DOI：
10.1007/s10544-019-0390-0
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发表时间：
2019
影响因子：
2.8
通讯作者：
Onesto V
中科院分区：
文献类型：
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作者：
Onesto V
在胚胎发生中，间充质凝聚是许多器官系统（包括软骨和骨）形成过程中的关键事件。在器官形成过程中，间充质细胞聚集并进行压实，同时激活发育程序。器官的最终三维形式以及细胞的命运可以受到形成冷凝物的大小和形状的影响。假设这一过程是间充质微环境中多尺度细胞相互作用的结果;然而，这些在体内研究是复杂的。重现关键表型的三维体外模型有助于我们理解调节这一基本发育过程的微环境相互作用。在这里，我们设计这样的模型，通过使用图像分析来指导设计的聚二甲基硅氧烷三维微结构作为细胞培养基板。这些微结构建立了几何约束的小鼠胚胎骨骼祖细胞的微团培养，影响凝聚的发展。我们首先确定关键的表型区分脸和肢芽微团文化的线性判别分析的形状描述符冷凝形态，这是用来指导合理设计的微图案化聚二甲基硅氧烷基板。高内涵的成像分析强调，微环境的几何形状影响冷凝的建立和增长。此外，细胞致力于在前5小时内建立凝聚;凝聚在17小时内达到其完整尺寸;之后它们增加细胞密度，同时保持尺寸至少7天。这些发现阐明了我们的模型在解剖间充质凝聚发展的关键方面的价值。
In embryogenesis, mesenchymal condensation is a critical event during the formation of many organ systems, including cartilage and bone. During organ formation, mesenchymal cells aggregate and undergo compaction while activating developmental programmes. The final three-dimensional form of the organ, as well as cell fates, can be influenced by the size and shape of the forming condensation. This process is hypothesized to result from multiscale cell interactions within mesenchymal microenvironments; however, these are complex to investigatein vivo. Three-dimensionalin vitromodels that recapitulate key phenotypes can contribute to our understanding of the microenvironment interactions regulating this fundamental developmental process. Here we devise such models by using image analysis to guide the design of polydimethylsiloxane 3D microstructures as cell culture substrates. These microstructures establish geometrically constrained micromass cultures of mouse embryonic skeletal progenitor cells which influence the development of condensations. We first identify key phenotypes differentiating face and limb bud micromass cultures by linear discriminant analysis of the shape descriptors for condensation morphology, which are used to guide the rational design of a micropatterned polydimethylsiloxane substrate. High-content imaging analysis highlights that the geometry of the microenvironment affects the establishment and growth of condensations. Further, cells commit to establish condensations within the first 5 h; condensations reach their full size within 17 h; following which they increase cell density while maintaining size for at least 7 days. These findings elucidate the value of our model in dissecting key aspects of mesenchymal condensation development.