Impaired Osteogenesis of Disease-Specific Induced Pluripotent Stem Cells Derived from a CFC Syndrome Patient
Abstract
:1. Introduction
2. Results
2.1. Generation and Characterization of CFC-iPSCs
2.2. Impaired Osteogenesis of MSCs Derived from CFC-iPSCs In Vitro
2.3. Enhanced ERK Signaling Activates SMAD2 Pathway in CFC-MSCs during Osteogenic Differentiation
2.4. Enhanced SMAD2 Pathway Downregulates SMAD1 Pathway in CFC-MSCs during Osteogenic Differentiation
2.5. Defective Osteogenesis in CFC-MSCs Is Ameliorated by Modulation of ERK, SMAD2, and SMAD1 Signaling Pathways
3. Discussion
4. Materials and Methods
4.1. Ethical Statement
4.2. Generation and Characterization of Human iPSCs
4.3. Differentiation of iPSCs into MSCs
4.4. FACS Analysis
4.5. Differentiation of MSCs into Osteoblasts, Chondroblasts, and Adipocytes
4.6. RT-PCR and Quantitative RT-PCR
4.7. Western Blotting
4.8. Transfection of OPN Small Interfering RNA
4.9. Chemicals and Growth Factors Used for Regulating Various Signaling Pathways
4.10. Statistical Analysis
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
CFC | Cardiofaciocutaneous |
ERK | Extracellular signal-regulated kinase |
iPSC | Induced pluripotent stem cell |
BRAF | Rapidly accelerated fibrosarcoma kinase B |
qRT-PCR | Quantitative real-time polymerase chain reaction |
MSC | Mesenchymal stem cell |
WT | Wild-type |
Ob | Osteoblast |
HA | Hydroxyapatite |
ALP | Alkaline phosphatase |
PPi | Pyrophosphate |
ENPP1 | Ectonucleotide pyrophasphatase/phosphodiesterase 1 |
ANKH | Ankylosis human |
OPN | Osteopontin |
BSP | Bone sialoprotein |
BMP | Bone morphogenetic protein |
TGF-β | Transforming growth factor-β |
RUNX2 | Runt-related transcription factor 2 |
OCN | Osteocalcin |
EB | Embryoid body |
ActA | Activin A |
SB | SB-431542 |
LDN | LDN-193189 |
PDGF | Platelet-derived growth factor-BB |
U | U0126 |
B4 | Bone morphogenic protein 4 |
si-OPN | Osteopontin small interfering RNA |
si-SCR | Scrambled small interfering RNA |
FACS | fluorescence-activated cell sorting |
α-MEM | α-minimum essential medium |
FBS | Fetal bovine serum |
SEM | Standard error of the mean |
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Choi, J.-Y.; Han, K.-M.; Kim, D.; Lee, B.-H.; Yoo, H.-W.; Choi, J.-H.; Han, Y.-M. Impaired Osteogenesis of Disease-Specific Induced Pluripotent Stem Cells Derived from a CFC Syndrome Patient. Int. J. Mol. Sci. 2017, 18, 2591. https://doi.org/10.3390/ijms18122591
Choi J-Y, Han K-M, Kim D, Lee B-H, Yoo H-W, Choi J-H, Han Y-M. Impaired Osteogenesis of Disease-Specific Induced Pluripotent Stem Cells Derived from a CFC Syndrome Patient. International Journal of Molecular Sciences. 2017; 18(12):2591. https://doi.org/10.3390/ijms18122591
Chicago/Turabian StyleChoi, Jung-Yun, Kyu-Min Han, Dongkyu Kim, Beom-Hee Lee, Han-Wook Yoo, Jin-Ho Choi, and Yong-Mahn Han. 2017. "Impaired Osteogenesis of Disease-Specific Induced Pluripotent Stem Cells Derived from a CFC Syndrome Patient" International Journal of Molecular Sciences 18, no. 12: 2591. https://doi.org/10.3390/ijms18122591