Viscoelastic microbead stress sensors and validation based on organoid mechanobiology

基于类器官力学生物学的粘弹性微珠应力传感器及验证

• 批准号: 467937258
• 负责人: Professor Dr.-Ing. Daniel Balzani
• 金额: --
• 依托单位: Institut Biofunktionelle Polymermaterialien
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/467937258?language=en
• 关键词: Viscoelastic; microbead; stress; sensors; validation

Mechanical stress exerted and experienced by cells plays an important role for tissue morphogenesis and organ formation. While techniques to quantify mechanical stresses are established, their study in living tissues and organisms remains challenging. Recently introduced hydrogel-based microbeads represent a new type of stress sensors. These deformable, spherical, cell-sized probes are injected into the intercellular space of cell aggregates or tissues and allow the direct interaction with neighboring cells. On the basis of the resulting deformations of these microbeads, stresses can be quantified at the cellular level. However, current sensors exhibit elastic material properties which, after introduction into a viscoelastic tissue environment, can influence the cellular behavior and thus lead to measurement artifacts. Furthermore, the rate-dependent interaction of the cells can not be captured. In addition to that, additional functionalization, e.g., regarding incorporation of fluorescent position and orientation markers, is hardly possible in available sensors. Therefore, we plan to develop novel microbead stress sensors in this project based on a viscoelastic hydrogel material which is highly flexible with regard to cell responsive functionalization. These microbeads will be produced by a microfluidic device and precisely characterized in terms of their mechanical properties at the micro- and macroscale. Furthermore, they will be incorporated into pluripotent stem cell-derived kidney organoids to explore their potential to quantify cellular stresses at different developmental stages of organoid growth. An already established method called “Computational Analysis of Cell Scale Stress Sensing” (COMPAX), which will be extended to the application of the viscoelastic stress sensors, will be used for the subsequent analysis of the microbeads. In this context, a new simulation setup for the stress calculations will be developed and validated. With this novel type of stress sensors, we expect an expansion of the range of mechanobiological tools as well as new spatiotemporal insights into the developmental processes during organoid formation enabling a better understanding of cellular processes.

细胞所承受的机械应力对组织形态发生和器官形成起着重要作用。虽然量化机械应力的技术已经建立，但它们在活组织和生物体中的研究仍然具有挑战性。最近推出的水凝胶微球代表了一种新型的应力传感器。这些可变形的球形细胞大小的探针被注入细胞聚集体或组织的细胞间空间，并允许与邻近细胞直接相互作用。根据这些微珠产生的变形，可以在细胞水平上量化应力。然而，电流传感器表现出弹性材料特性，在引入粘弹性组织环境后，可能会影响细胞行为，从而导致测量伪影。此外，不能捕捉到细胞之间依赖于速率的相互作用。除此之外，在现有的传感器中几乎不可能实现额外的功能化，例如关于加入荧光位置和方向标记。因此，我们计划在这个项目中开发基于粘弹性水凝胶材料的新型微珠应力传感器，这种材料在细胞响应功能化方面具有高度的灵活性。这些微珠将由微流控装置生产，并在微观和宏观尺度上精确表征其机械性能。此外，它们将被整合到多能干细胞来源的肾脏器官中，以探索它们在器官生长的不同发育阶段量化细胞压力的潜力。一种已经建立的名为“细胞尺度应力传感的计算分析”(COMPAX)的方法将被用于随后的微珠分析，该方法将扩展到粘弹性应力传感器的应用。在这种情况下，将开发和验证用于应力计算的新的模拟设置。有了这种新型的应力传感器，我们期待着机械生物学工具的范围扩大，以及对有机体形成过程中的发育过程的新的时空洞察，从而能够更好地理解细胞过程。