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Abstract：
3D cell culture has emerged as a promising approach to replicate the complex behaviors of cells within living organisms. This study aims to analyze spatiotemporal behavior of the morphological characteristics of cell structure at multiscale in 3D scaffold-free spheroids using chondrogenic progenitor ATDC5 cells. Over a 14-day culture period, it exhibited cell hypertrophy in the spheroids regarding cellular and nuclear size as well as changes in morphology. Moreover, biological analysis indicated a signification up-regulation of normal chondrocyte as well as hypertrophic chondrocyte markers, suggesting early hypertrophic chondrocyte differentiation. Cell nuclei underwent changes in volume, sphericity, and distribution in spheroid over time, indicating alterations in chromatin organization. The ratio of chromatin condensation volume to cell nuclear volume decreased as the cell nuclei enlarged, potentially signifying changes in chromatin state during hypertrophic chondrocyte differentiation. Our image analysis techniques in this present study enabled detailed morphological measurement of cell structure at multi-scale, which can be applied to various 3D culture models for in-depth investigation.
摘要：
3D细胞培养已经成为一种有希望的方法来复制活生物体内细胞的复杂行为。本研究旨在分析使用软骨形成祖细胞ATDC5细胞在3D无支架球体中的多尺度细胞结构形态特征的时空行为。在14天的文化期内，在细胞和核大小以及形态变化方面，它在球状体中表现出细胞肥大。此外，生物学分析表明正常软骨细胞和肥大软骨细胞标志物有明显的上调，提示早期肥大软骨细胞分化。细胞核经历了体积的变化，球形，随着时间的推移在球体中的分布，表明染色质组织的改变。染色质浓缩体积与细胞核体积的比率随着细胞核的增大而降低，肥大软骨细胞分化过程中染色质状态的潜在变化。在本研究中，我们的图像分析技术能够在多尺度下对细胞结构进行详细的形态学测量，可以应用于各种3D培养模型进行深入研究。
