Zhang Qi, Deng Shuwen, Sun Ke, Lin Shiyu, Lin Yunfeng, Zhu Bofeng, Cai Xiaoxiao
State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China.
Department of Stomatology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China.
Cell Prolif. 2017 Dec;50(6). doi: 10.1111/cpr.12385. Epub 2017 Sep 18.
The crosstalk between chondrocytes and adipose-derived stem cells (ADSCs) could regulate the secretion of multiple growth factors. However, it is not clear how the paracrine action in co-culture systems affect cell migration. This study focused on the changes of cell migration of ADSCs and chondrocytes in co-culture conditions.
Primary ADSCs and chondrocytes were isolated from Sprague-Dawley rat. Transwell co-culture systems, inoculated with ADSCs and chondrocytes, were established in vitro. The morphology of the cells was observed 7 days post-seeding by inverted phase-contrast microscope. Additionally, the cytoskeleton changes were investigated by immunofluorescence staining. To detect the abundance of Vinculin, we used immunofluorescence and Western blotting. Additionally, the expression level of MMP-2, Hey1 and Hes1 was examined to determine the mechanisms of co-culture-induced cell migration changes.
The migration of ADSCs and chondrocytes in co-culture conditions significantly decreased compared with that in mono-culture groups, accompanied by the decrease of filopodia and the expression level of MMP-2.
The overall study showed that the migration of ADSCs and chondrocytes differs significantly depending on culture conditions. Moreover, the Notch signalling pathway may be involved in this process. Accordingly, by studying changes in migration caused by co-culture, we obtained new insight into the crosstalk between ADSCs and chondrocytes.
软骨细胞与脂肪来源干细胞(ADSCs)之间的相互作用可调节多种生长因子的分泌。然而,共培养体系中的旁分泌作用如何影响细胞迁移尚不清楚。本研究聚焦于共培养条件下ADSCs和软骨细胞的细胞迁移变化。
从Sprague-Dawley大鼠分离原代ADSCs和软骨细胞。在体外建立接种ADSCs和软骨细胞的Transwell共培养体系。接种7天后,用倒置相差显微镜观察细胞形态。此外,通过免疫荧光染色研究细胞骨架变化。为检测纽蛋白的丰度,我们使用了免疫荧光和蛋白质印迹法。此外,检测MMP-2、Hey1和Hes1的表达水平,以确定共培养诱导的细胞迁移变化的机制。
与单培养组相比,共培养条件下ADSCs和软骨细胞的迁移显著降低,同时伴有丝状伪足减少和MMP-2表达水平降低。
总体研究表明,ADSCs和软骨细胞的迁移因培养条件而异。此外,Notch信号通路可能参与这一过程。因此,通过研究共培养引起的迁移变化,我们对ADSCs和软骨细胞之间的相互作用有了新的认识。