Department of Mechanical Systems Engineering, Graduate School of Engineering, Nagoya University, Nagoya, Japan.
Department of Biosystems Science, Institute for Life and Medical Sciences, Kyoto University, Kyoto, Japan.
Cell Tissue Res. 2024 Sep;397(3):263-274. doi: 10.1007/s00441-024-03905-7. Epub 2024 Jul 23.
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.
3D 细胞培养已成为一种有前途的方法，可以复制活生物体中细胞的复杂行为。本研究旨在使用软骨祖细胞 ATDC5 细胞分析无支架 3D 球体中细胞结构形态特征的时空行为。在 14 天的培养过程中，细胞在球体中发生肥大，表现为细胞和核大小以及形态的变化。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的显著上调，表明早期肥大软骨细胞分化。随着时间的推移，细胞核的体积、球形度和分布在球体中发生变化，表明染色质组织发生改变。随着细胞核的增大，染色质浓缩体积与细胞核体积的比值降低，这可能表明在肥大软骨细胞分化过程中染色质状态发生了变化。本研究中的图像分析技术能够对多尺度的细胞结构进行详细的形态学测量，可应用于各种 3D 培养模型进行深入研究。
