Osteoblasts Derived from Induced Pluripotent Stem Cells form Calcified Structures in Scaffolds Both In Vitro and In Vivo

Abstract

Reprogramming somatic cells into an embryonic stem (ES) cell-like state, or induced pluripotent stem (iPS) cells, has emerged as a promising new venue for customized cell therapies. In this study, we performed directed differentiation to assess the ability of murine iPS cells to differentiate into bone, cartilage and fat in vitro and to maintain an osteoblast phenotype on a scaffold in vitro and in vivo. Embryoid bodies derived from murine iPS cells were cultured in differentiation medium for eight to twelve weeks. Differentiation was assessed by lineage specific morphology, gene expression, histological stain and immunostaining to detect matrix deposition. After 12 weeks of expansion, iPS derived osteoblasts were seeded in a gelfoam matrix followed by subcutaneous implantation in syngenic ICR mice. Implants were harvested at 12 weeks, and histological analyses of cell, mineral and matrix content were performed. Differentiation of iPS cells into mesenchymal lineages of bone, cartilage and fat was confirmed by morphology, and expression of lineage specific genes. Isolated implants of iPS cell derived osteoblasts expressed matrices characteristic of bone, including osteocalcin and bone sialoprotein. Implants were also stained with alizarin red and von Kossa, demonstrating mineralization and persistence of an osteoblast phenotype. Recruitment of vasculature and microvascularization of the implant was also detected. Taken together, these data demonstrate functional osteoblast differentiation from iPS cells both in vitro and in vivo and reveal a source of cells which merit evaluation for their potential uses in orthopaedic medicine and understanding of molecular mechanisms of orthopaedic disease.

Bioengineering of Articular Cartilage: Past, Present and Future

“…Recently, induced pluripotent stem cells (iPSCs) have been used to different both osteogenic and chondrogenic cell types [58,59]. Like hESCs, iPSCs has the potential to provide great scope for cellular expansion and differentiation compared to mesenchymal stem cells, without the same ethical problems [60].…”

Differentiation behavior of iPS cells cultured on PLGA with osteoinduction medium

“…But the method of cell and scaffold attachment, iPS-MSCs differentiation must be confirmed with xeno-free. And the application of iPS cells and scaffold will require much more preclinical evaluation and testing, including safety assessment and a safer approach as it is in other aspects 35,36) . In the near feature, when osteogenesis in three-dimensional scaffolds and the safety in vivo are confirmed, we may be able to make a clinical jawbone reconstruction being the same shape as the excised bone, reproduced by a threedimensional printer.…”

Potential for pharmacological manipulation of human embryonic stem cells

“…Such diseases/ disorders include Parkinson's disease (Chung et al, 2011;Kriks et al, 2011;Kim et al, 2011b), retinal degeneration (Tucker et al, 2011), spinal chord injury (Nori et al, 2011), hypopigmentation disorders (Nissan et al, 2011), Alzheimer's disease (Bissonnette et al, 2011) and orthopaedic disease (Bilousova et al, 2011), and efficient protocols for derivation of specific cell types from hESC and hiPSCs may lead to the use of such cells to treat human disease. To this end, multiple small molecule drugs that can modulate the differentiation of clinical-grade hESCs (Ilic et al, 2011) or hiPSCs have been discovered and may be used in the future in cGMP-compliant differentiation protocols to produce transplantable cells.…”