Differentiating Chondrocytes from Peripheral Blood-derived Human Induced Pluripotent Stem Cells

作者：Li, Yueying^*; Hai, Yong; Chen, Jiayu; Liu, Tie

来源：Jove-Journal of Visualized Experiments, 2017, (125): e55722.

DOI：10.3791/55722

• Developmental Biology
• Issue 125
• induced pluripotent stem cells
• peripheral blood
• differentiation
• embryoid body
• fibroblastic cells
• chondrocyte

In this study, we used peripheral blood cells (PBCs) as seed cells to produce chondrocytes via induced pluripotent stem cells (iPSCs) in an integration-free method. Following embryoid body (EB) formation and fibroblastic cell expansion, the iPSCs are induced for chondrogenic differentiation for 21 days under serum-free and xeno-free conditions. After chondrocyte induction, the phenotypes of the cells are evaluated by morphological, immunohistochemical, and biochemical analyses, as well as by the quantitative real-time PCR examination of chondrogenic differentiation markers. The chondrogenic pellets show positive alcian blue and toluidine blue staining. The immunohistochemistry of collagen II and X staining is also positive. The sulfated glycosaminoglycan (sGAG) content and the chondrogenic differentiation markers COLLAGEN 2 (COL2), COLLAGEN 10 (COL10), SOX9, and AGGRECAN are significantly upregulated in chondrogenic pellets compared to hiPSCs and fibroblastic cells. These results suggest that PBCs can be used as seed cells to generate iPSCs for cartilage repair, which is patient-specific and cost-effective.

• 出版日期2017-7
• 单位同济大学; 中国科学院北京基因组研究所; 首都医科大学