In Vitro Fabrication of Hybrid Bone/Cartilage Complex Using Mouse Induced Pluripotent Stem Cells
Abstract
:1. Introduction
2. Results
2.1. Effects of Osteogenic (Os) and Osteo-Chondrogenic (Os-Chon) Induction on Osteochondrogenic Differentiation of 3D-iPSC Constructs
2.1.1. Expression of Osteogenic Marker Genes
2.1.2. Expression of Chondrogenic Marker Genes
2.1.3. Expression of Pluripotency Marker Genes
2.2. Effects of Os Induction on Mineralization and Cartilage Formation in Osteogenically Induced iPSC (OI-iPSC) Constructs
2.3. Effects of Os-Chon Induction on Mineralization and Cartilage Formation in Osteo-Chondrogenically Induced iPSC (OCI-iPSC) Constructs
2.4. Effects of Os and Os-Chon Induction on the Ratio of Calcification and Cartilage Formation in 3D-iPSC Constructs
2.5. Effects of Os and Os-Chon Induction on Mesodermal Lineage Commitment in 3D-iPSC Constructs
2.5.1. Early Mesodermal Marker Expression
2.5.2. Paraxial Mesoderm (PM) Marker Genes
2.5.3. Lateral Plate Mesoderm (LM) Marker Genes
3. Discussion
4. Materials and Methods
4.1. iPSC Culture
4.2. Fabrication of 3D-iPSC Spheres
4.3. Osteogenic Induction of 3D-iPSC Constructs
4.4. Osteochondrogenic Induction of 3D-iPSC Constructs
4.5. Reverse Transcription Polymerase Chain Reaction (RT-PCR) Analysis
4.6. Histochemical Analyses
4.7. Histomorphometric Quantification
4.8. Immunohistochemical Staining
4.9. Statistical Analyses
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Limraksasin, P.; Kondo, T.; Zhang, M.; Okawa, H.; Osathanon, T.; Pavasant, P.; Egusa, H. In Vitro Fabrication of Hybrid Bone/Cartilage Complex Using Mouse Induced Pluripotent Stem Cells. Int. J. Mol. Sci. 2020, 21, 581. https://doi.org/10.3390/ijms21020581
Limraksasin P, Kondo T, Zhang M, Okawa H, Osathanon T, Pavasant P, Egusa H. In Vitro Fabrication of Hybrid Bone/Cartilage Complex Using Mouse Induced Pluripotent Stem Cells. International Journal of Molecular Sciences. 2020; 21(2):581. https://doi.org/10.3390/ijms21020581
Chicago/Turabian StyleLimraksasin, Phoonsuk, Takeru Kondo, Maolin Zhang, Hiroko Okawa, Thanaphum Osathanon, Prasit Pavasant, and Hiroshi Egusa. 2020. "In Vitro Fabrication of Hybrid Bone/Cartilage Complex Using Mouse Induced Pluripotent Stem Cells" International Journal of Molecular Sciences 21, no. 2: 581. https://doi.org/10.3390/ijms21020581