In Vivo 3D Histomorphometry Quantifies Bone Apposition and Skeletal Progenitor Cell Differentiation.
In Vivo 3D Histomorphometry Quantifies Bone Apposition and Skeletal Progenitor Cell Differentiation.
复制标题
体内 3D 组织形态计量学可量化骨沉积和骨骼祖细胞分化。
DOI：
10.1038/s41598-018-23785-6
复制
发表时间：
2018
影响因子：
4.6
通讯作者：
Intini,Giuseppe
中科院分区：
文献类型：
--
作者：
Yeh,Shu-ChiA;Wilk,Katarzyna;Lin,CharlesP;Intini,Giuseppe
组织形态计量学和Micro-CT常用于评估骨重建和骨微结构。这些方法通常需要单独的动物队列来分析3D形态学变化，并涉及耗时的免疫组织化学制备。活体显微镜（IVM）结合小鼠遗传学可能是一个有吸引力的选择，以获得骨结构的测量，同时进行纵向监测的动态细胞processesin体内。在这项研究中，我们利用双光子，荧光IVM与谱系追踪报告小鼠模型的图像骨骼干细胞（SSC）在他们的颅骨缝龛和分析其分化后的命运与经典的Wnt通路（重组Wnt 3a）的激动剂刺激。骨形成、缝合体积和细胞动力学的Ourin活体组织形态计量学分析表明，重组Wnt 3a诱导新骨形成、分化和SSC后代并入新形成的骨中。因此，IVM技术可以为传统的静态2D组织形态测量提供额外的动态3D信息。
Histomorphometry and Micro-CT are commonly used to assess bone remodeling and bone microarchitecture. These approaches typically require separate cohorts of animals to analyze 3D morphological changes and involve time-consuming immunohistochemistry preparation. Intravital Microscopy (IVM) in combination with mouse genetics may represent an attractive option to obtain bone architectural measurements while performing longitudinal monitoring of dynamic cellular processesin vivo. In this study we utilized two-photon, multicolor fluorescence IVM together with a lineage tracing reporter mouse model to image skeletal stem cells (SSCs) in their calvarial suture niche and analyze their differentiation fate after stimulation with an agonist of the canonical Wnt pathway (recombinant Wnt3a). Ourin vivohistomorphometry analyses of bone formation, suture volume, and cellular dynamics showed that recombinant Wnt3a induces new bone formation, differentiation and incorporation of SSCs progeny into newly forming bone. IVM technology can therefore provide additional dynamic 3D information to the traditional static 2D histomorphometry.