Comparative Analysis of Mouse-Induced Pluripotent Stem Cells and Mesenchymal Stem Cells During Osteogenic Differentiation In Vitro

Abstract

Induced pluripotent stem cells (iPSCs) can differentiate into mineralizing cells and are, therefore, expected to be useful for bone regenerative medicine; however, the characteristics of iPSC-derived osteogenic cells remain unclear. Here, we provide a direct in vitro comparison of the osteogenic differentiation process in mesenchymal stem cells (MSCs) and iPSCs from adult C57BL/6J mice. After 30 days of culture in osteogenic medium, both MSCs and iPSCs produced robustly mineralized bone nodules that contained abundant calcium phosphate with hydroxyapatite crystal formation. Mineral deposition was significantly higher in iPSC cultures than in MSC cultures. Scanning electron microscopy revealed budding matrix vesicles in early osteogenic iPSCs; subsequently, the vesicles propagated to exhibit robust mineralization without rich fibrous structures. Early osteogenic MSCs showed deposition of many matrix vesicles in abundant collagen fibrils that became solid mineralized structures. Both cell types demonstrated increased expression of osteogenic marker genes, such as runx2, osterix, dlx5, bone sialoprotein (BSP), and osteocalcin, during osteogenesis; however, real-time reverse transcription-polymerase chain reaction array analysis revealed that osteogenesis-related genes encoding mineralization-associated molecules, bone morphogenetic proteins, and extracellular matrix collagens were differentially expressed between iPSCs and MSCs. These data suggest that iPSCs are capable of differentiation into mature osteoblasts whose associated hydroxyapatite has a crystal structure similar to that of MSCassociated hydroxyapatite; however, the transcriptional differences between iPSCs and MSCs could result in differences in the mineral and matrix environments of the bone nodules. Determining the biological mechanisms underlying cell-specific differences in mineralization during in vitro iPSC osteogenesis may facilitate the development of clinically effective engineered bone.

Notch signaling partly regulates the osteogenic differentiation of retinoic acid-treated murine induced pluripotent stem cells

“…The products were electrophoresed in a 1.8% agarose gel and stained with ethidium bromide. The primer sequences are shown in Table 1 (4,(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28) Statistical analysis Data are shown as mean ± standard deviation. Statistical analysis was performed using the two-independent samples t-test for two-group comparisons and one-way …”

“…Murine induced pluripotent stem cells (miPSCs) are currently considered an attractive cell type for osteogenic differentiation (2). For the efficient osteogenic differentiation of mGF-iPSCs, all-trans retinoic acid (RA)-treated embryoid bodies, which contain immature mesenchymal cells, are used (3,4). Osteogenic induction markedly promotes osteoblastrelated gene expression and mineral deposition by these cells.…”

“…Osteogenic induction markedly promotes osteoblastrelated gene expression and mineral deposition by these cells. However, it should be noted that the osteogenesisrelated genes expressed by mGF-iPSCs differ from those expressed by mesenchymal stem cells (MSCs), implying that different regulatory mechanisms are involved (4). mGF-iPSCs may be an excellent cell source for bone regeneration treatments.…”

Feeder Cell Sources and Feeder-Free Methods for Human iPS Cell Culture

“…In contrast to the case with mouse ESCs, LIF does not satisfactorily support the self-renewal of human ESCs [4]. Nonetheless, SNL76/7 cells basically inherit the nature of STO cells; therefore, SNL 76/7 cells can be used as feeder cells for both mouse [6,30,31] and human [1,28,30] ESCs/iPSCs. Bradley also established a puromycin-resistant derivative of SNL76/7 cells (SNLP76/7-4 cell line: Fig.…”

Patient-Derived Human Induced Pluripotent Stem Cells From Gingival Fibroblasts Composited With Defined Nanohydroxyapatite/Chitosan/Gelatin Porous Scaffolds as Potential Bone Graft Substitutes

“…The development of induced pluripotent stem cells (iPSCs) was a milestone for stem cell research, not only because of their unlimited self-renewal capacity and pluripotent differentiation possibility, but also because iPSCs are derived autologously from patient somatic cells, which could avoid ethical problems. Therefore, iPSCs have been a promising cell source for clinical use in bone regenerative medicine [10,11].…”