期刊
JOURNAL OF TISSUE ENGINEERING
卷 13, 期 -, 页码 -出版社
SAGE PUBLICATIONS INC
DOI: 10.1177/20417314221114616
关键词
BMP-4; cartilage regeneration; iPS cells; shaking culture; transcriptional activation
资金
- Japan Society for the Promotion of Science [JP16K20480, JP16K15797, JP16H05519, JP19H03840]
In this study, a new iPSC-Tet/BMP-4 line was established and an efficient approach combining transcriptional activation of BMP-4 and a 3D shaking suspension culture system was developed to differentiate iPSCs into chondrocytes. The culture system could be a useful tool for further investigation of the mechanism of BMP-4 in regulating iPSC differentiation towards the chondrogenic lineage, and should facilitate research in cartilage development, repair, and osteoarthritis.
Induced pluripotent stem cells (iPSCs) offer an unlimited source for cartilage regeneration as they can generate a wide spectrum of cell types. Here, we established a tetracycline (tet) controlled bone morphogenetic protein-4 (BMP-4) expressing iPSC (iPSC-Tet/BMP-4) line in which transcriptional activation of BMP-4 was associated with enhanced chondrogenesis. Moreover, we developed an efficient and simple approach for directly guiding iPSC-Tet/BMP-4 differentiation into chondrocytes in scaffold-free cartilaginous pellets using a combination of transcriptional activation of BMP-4 and a 3D shaking suspension culture system. In chondrogenic induction medium, shaking culture alone significantly upregulated the chondrogenic markers Sox9, Col2a1, and Aggrecan in iPSCs-Tet/BMP-4 by day 21. Of note, transcriptional activation of BMP-4 by addition of tet (doxycycline) greatly enhanced the expression of these genes. The cartilaginous pellets derived from iPSCs-Tet/BMP-4 showed an oval morphology and white smooth appearance by day 21. After day 21, the cells presented a typical round morphology and the extracellular matrix was stained intensively with Safranin O, alcian blue, and type II collagen. In addition, the homogenous cartilaginous pellets derived from iPSCs-Tet/BMP-4 with 28 days of induction repaired joint osteochondral defects in immunosuppressed rats and integrated well with the adjacent host cartilage. The regenerated cartilage expressed the neomycin resistance gene, indicating that the newly formed cartilage was generated by the transplanted iPSCs-Tet/BMP-4. Thus, our culture system could be a useful tool for further investigation of the mechanism of BMP-4 in regulating iPSC differentiation toward the chondrogenic lineage, and should facilitate research in cartilage development, repair, and osteoarthritis.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据