Laboratory of Cell and Molecular Mechanisms of Histogenesis, Kol'tsov Institute of Developmental Biology, Russian Academy of Sciences, Moscow, Russia.
Methods Mol Biol. 2022;2520:59-79. doi: 10.1007/7651_2021_405.
The 3D embryoid body (EB) differentiation model is a promising tool for fundamental cell biology and drug discovery studies assessing the compound effects on mammalian and human development. This 3D cell model allows for analyzing spatiotemporal changes during morphogenesis and differentiation. A combination of confocal microscopy with high content image analysis (HCIA) can significantly improve the study of spatiotemporal patterns of early embryonic lineages and compound efficacy and toxicity testing by enhancing the identification and quantification of various cell types. HCIA can be used to assess the EB architecture through quantitative and qualitative characteristics, such as viability and apoptosis, identification, localization, ratio and timing for various types of early embryonic cells, dimensions of compartments of proliferating and differentiating cells, changes in the size and shape of EBs, and translocation of individual cells and cell layers. This chapter describes a comprehensive framework for HCIA for 3D EB differentiation model that allows investigators to analyze EB growth, differentiation, and morphogenetic dynamics.
三维胚胎体 (EB) 分化模型是一种很有前途的工具，可用于基础细胞生物学和药物发现研究，评估化合物对哺乳动物和人类发育的复合影响。这种 3D 细胞模型允许分析形态发生和分化过程中的时空变化。共聚焦显微镜与高内涵图像分析 (HCIA) 的结合可以通过增强对各种细胞类型的识别和定量，显著改善早期胚胎谱系和化合物功效和毒性测试的时空模式研究。HCIA 可用于通过定量和定性特征评估 EB 结构，例如活力和凋亡、各种早期胚胎细胞的鉴定、定位、比例和时间、增殖和分化细胞区室的尺寸、EB 大小和形状的变化，以及单个细胞和细胞层的易位。本章描述了一个用于 3D EB 分化模型的 HCIA 的综合框架，允许研究人员分析 EB 的生长、分化和形态发生动力学。
