Effects of Cytoskeleton Modulation on the Phenotype of Articular Chondrocytes in Monolayer and Pellet Culture
Effects of Cytoskeleton Modulation on the Phenotype of Articular Chondrocytes in Monolayer and Pellet Culture
复制标题
细胞骨架调节对单层和颗粒培养关节软骨细胞表型的影响
DOI：
10.1166/jbt.2015.1397
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发表时间：
2015-12-01
影响因子：
0.1
通讯作者：
Yang, Liu
中科院分区：
文献类型：
--
作者：
Chen, Cheng;Yin, Li;Yang, Liu
以前的研究表明，软骨细胞在二维培养能够保持软骨细胞表型时，肌动蛋白应力纤维的形成是化学抑制细胞松弛素;软骨细胞在三维培养通常表达更好的软骨形成表型比在二维，因为肌动蛋白应力纤维的形成是物理抑制细胞聚集，而不是细胞扩散。除了肌动蛋白，另一种细胞骨架蛋白波形蛋白也被证明影响软骨细胞表型。这些发现表明软骨细胞的成软骨表达与细胞骨架组织有关。为了系统地研究细胞骨架调控对软骨细胞表型的影响，本研究采用二维单层和三维颗粒培养兔关节软骨细胞，分别破坏波形蛋白、微管蛋白和肌动蛋白网络，并检测软骨细胞表型的变化。我们发现，波形蛋白中断诱导减少II型胶原和聚集蛋白聚糖，并在3D软骨细胞的程度大于在2D中;微管蛋白中断显示软骨细胞表型的影响不大;肌动蛋白中断只促进软骨细胞在2D中II型胶原的表达，而不是在3D中。我们的研究结果表明，波形蛋白在维持软骨细胞表型的关键作用，并进一步提供了对体外表型操纵的见解。
Previous studies have shown that chondrocytes in 2D culture are capable of keeping chondrocytic phenotype when formation of actin stress fiber is chemically restrained by cytochalasin; chondrocytes in 3D culture normally express better chondrogenic phenotype than in 2D, because formation of actin stress fiber is physically inhibited by cell aggregation instead of cell spreading. Besides actin, another cytoskeletal protein vimentin has also been shown to affect chondrocyte phenotype. These findings indicate that the chondrogenic expression of chondrocytes is associated with cytoskeleton organization. To systematically investigate the effects of cytoskeleton modulation on chondrocyte phenotype, here we cultured rabbit articular chondrocytes in 2D monolayer and 3D pellet, disrupted vimentin, tubulin, or actin network respectively, and measured the changes of chondrocyte phenotype. We found that vimentin disruption induced a reduction of type II collagen and aggrecan, and the extent was greater for chondrocytes in 3D than in 2D; tubulin disruption showed little effects on chondrocyte phenotype; actin disruption only promoted the expression of type II collagen for chondrocytes in 2D not in 3D. Our findings suggest a critical role of vimentin in the maintenance of chondrocyte phenotype, and further provide insights toward in vitro phenotype manipulation.