Imaging and detecting intercellular tensile forces in spheroids and embryoid bodies using lipid-modified DNA probes.
Imaging and detecting intercellular tensile forces in spheroids and embryoid bodies using lipid-modified DNA probes.
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DOI：
10.3389/fcell.2023.1220079
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发表时间：
2023
影响因子：
5.5
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中科院分区：
文献类型：
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细胞不断地感受并对用于调节其生理和功能的不同物理力作出反应。我们测量这些力学信号的能力对于理解各种机械感知和机械转导过程的基础至关重要。虽然已经开发了多种策略来研究二维（2D）细胞培养单层内的机械力，但在真实的三维（3D）细胞模型中细胞 - 细胞连接处的力测量仍然相当罕见。考虑到在真实的生物系统中，细胞受到来自三维方向的力，因此在其原生环境中测量这些分子力对于更好地理解不同的发育和疾病过程至关重要。我们最近开发了一种基于DNA的分子探针，用于测量2D细胞模型中的细胞间张力。在此，我们将报告这些分子张力探针的进一步开发以及首次用于可视化和检测3D球体和类胚体（EBs）内的机械力。这些探针可以通过连接的脂质部分自发地锚定在活细胞膜上。通过改变这些DNA探针的浓度及其孵育时间，我们首先表征了探针穿透并加载到不同大小的肿瘤球体和干细胞类胚体上的动力学和效率。经过优化，我们首次进一步对这些3D球体和类胚体中E - 钙黏蛋白介导的力进行了成像和测量。我们的结果表明，这些基于DNA的分子张力探针可用于研究目标机械转导过程的时空分布。这些强大的成像工具可能有潜在应用，以填补二维系统中生物力学的现有研究与真实三维细胞复合体中的研究之间的差距。
Cells continuously experience and respond to different physical forces that are used to regulate their physiology and functions. Our ability to measure these mechanical cues is essential for understanding the bases of various mechanosensing and mechanotransduction processes. While multiple strategies have been developed to study mechanical forces within two-dimensional (2D) cell culture monolayers, the force measurement at cell-cell junctions in real three-dimensional (3D) cell models is still pretty rare. Considering that in real biological systems, cells are exposed to forces from 3D directions, measuring these molecular forces in their native environment is thus highly critical for the better understanding of different development and disease processes. We have recently developed a type of DNA-based molecular probe for measuring intercellular tensile forces in 2D cell models. Herein, we will report the further development and first-time usage of these molecular tension probes to visualize and detect mechanical forces within 3D spheroids and embryoid bodies (EBs). These probes can spontaneously anchor onto live cell membranes via the attached lipid moieties. By varying the concentrations of these DNA probes and their incubation time, we have first characterized the kinetics and efficiency of probe penetration and loading onto tumor spheroids and stem cell EBs of different sizes. After optimization, we have further imaged and measured E-cadherin-mediated forces in these 3D spheroids and EBs for the first time. Our results indicated that these DNA-based molecular tension probes can be used to study the spatiotemporal distributions of target mechanotransduction processes. These powerful imaging tools may be potentially applied to fill the gap between ongoing research of biomechanics in 2D systems and that in real 3D cell complexes.