Key Laboratory of Biological Medicines in Universities of Shandong Province, Weifang Medical University, Weifang, 261053, China.
Key Laboratory of Biological Medicines in Universities of Shandong Province, Weifang Medical University, Weifang, 261053, China.
Biomed Pharmacother. 2019 Sep;117:109146. doi: 10.1016/j.biopha.2019.109146. Epub 2019 Jul 2.
Mesenchymal stem cells and chondrocytes are an important source of the cells for cartilage tissue engineering. Therefore, the culture and expansion methods of these cells need to be improved to overcome the aging of chondrocytes and induced chondrogenic differentiation of mesenchymal stem cells. The aim of this study was to expand the cells for cartilage tissue engineering by combining the advantages of growing cells in co-culture and under a mechanically-stimulated environment. Rabbit chondrocytes and co-cultured cells (bone mesenchymal stem cells and chondrocytes) were subjected to cyclic sinusoidal dynamic tensile mechanical stimulationusing the FX-4000 tension system. Chondrocyte proliferation was assayed by flow cytometry and CFSE labeling. The cell cartilage phenotype was determined by detecting GAG, collagen II and TGF-β1 protein expression by ELISA and the Col2α1, TGF-β1 and Sox9 gene expression by RT-PCR. The results show that the co-culture improved both the proliferation ability of chondrocytes and the cartilage phenotype of co-cultured cells. A proper cyclic sinusoidal dynamic tensile mechanical stimulation improved the proliferation ability and cartilage phenotype of chondrocytes and co-cultured cells. These results suggest that the co-culture of mesenchymal stem cells with chondrocytes and proper mechanical stimulation may be an appropriate way to rapidly expand the cells that have an improved cartilage phenotype for cartilage tissue engineering.
间充质干细胞和软骨细胞是软骨组织工程细胞的重要来源。因此，需要改进这些细胞的培养和扩增方法，以克服软骨细胞的衰老和诱导间充质干细胞的软骨分化。本研究旨在通过结合细胞共培养和机械刺激环境下生长细胞的优势，来扩增用于软骨组织工程的细胞。使用 FX-4000 张力系统对兔软骨细胞和共培养细胞（骨间充质干细胞和软骨细胞）进行周期性正弦动态拉伸机械刺激。通过流式细胞术和 CFSE 标记法检测细胞增殖。通过 ELISA 检测 GAG、胶原 II 和 TGF-β1 蛋白表达，通过 RT-PCR 检测 Col2α1、TGF-β1 和 Sox9 基因表达，来确定细胞的软骨表型。结果表明，共培养提高了软骨细胞的增殖能力和共培养细胞的软骨表型。适当的周期性正弦动态拉伸机械刺激提高了软骨细胞和共培养细胞的增殖能力和软骨表型。这些结果表明，间充质干细胞与软骨细胞的共培养和适当的机械刺激可能是一种快速扩增具有改善软骨表型的细胞用于软骨组织工程的合适方法。
