Directed Differentiation of Human Induced Pluripotent Stem Cells Toward Bone and Cartilage: In Vitro Versus In Vivo Assays

The ability to generate large numbers of bone‐ and cartilage‐forming cells from induced pluripotent stem cells (iPSCs) would mark a major advance in tissue engineering. A number of protocols exist, but the overall quality and consistency of this type of differentiation are under‐reported. In this study, the authors analyzed differentiated iPSCs in vitro and in vivo by stringent criteria, and found that in vitro analysis does not predict in vivo differentiation. The ability to differentiate induced pluripotent stem cells (iPSCs) into committed skeletal progenitors could allow for an unlimited autologous supply of such cells for therapeutic uses; therefore, we attempted to create novel bone‐forming cells from human iPSCs using lines from two distinct tissue sources and methods of differentiation that we previously devised for osteogenic differentiation of human embryonic stem cells, and as suggested by other publications. The resulting cells were assayed using in vitro methods, and the results were compared with those obtained from in vivo transplantation assays. Our results show that true bone was formed in vivo by derivatives of several iPSC lines, but that the successful cell lines and differentiation methodologies were not predicted by the results of the in vitro assays. In addition, bone was formed equally well from iPSCs originating from skin or bone marrow stromal cells (also known as bone marrow‐derived mesenchymal stem cells), suggesting that the iPSCs did not retain a “memory” of their previous life. Furthermore, one of the iPSC‐derived cell lines formed verifiable cartilage in vivo, which likewise was not predicted by in vitro assays.