Cytoskeleton systems, actin microfilaments, microtubules (MTs) and intermediate filaments (IFs) provide the biomechanical stability and spatial organization in cells. To understand the specific contributions of each cytoskeleton systems to intrinsic properties of spheroids, we've scrutinized the effects of the cytoskeleton perturbants, cytochalasin D (Cyto D), nocodazole (Noc) and withaferin A (WFA) on fusion, spreading on adhesive surface, morphology and biomechanics of chondrospheres (CSs). We confirmed that treatment with Cyto D but not with Noc or WFA severely affected CSs fusion and spreading dynamics and significantly reduced biomechanical properties of cell aggregates. Noc treatment affected spheroids spreading but not the fusion and surprisingly enhanced their stiffness. Vimentin intermediate filaments (VIFs) reorganization affected CSs spreading only. The analysis of all three cytoskeleton systems contribution to spheroids intrinsic properties was performed for the first time.

细胞骨架系统，肌动蛋白微丝，微管（MTs）和中间丝（IFs）在细胞中提供生物力学稳定性和空间组织。为了了解每个细胞骨架系统对球体固有特性的特定贡献，我们仔细研究了细胞骨架扰动剂，细胞松弛素D（Cyto D），诺考达唑（Noc）和Withaferin A（WFA）对融合的影响，并扩散到胶粘剂表面，软骨的形态和生物力学。我们证实，用Cyto D处理但不使用Noc或WFA处理会严重影响CS融合和扩散动力学，并显着降低细胞聚集体的生物力学性能。Noc处理影响球状体的扩散，但不影响融合，并且出人意料地增强了球状体的刚度。波形蛋白中间丝（VIF）重组仅影响CS的扩散。首次进行了对所有三个细胞骨架系统对球体内在特性贡献的分析。