Intermittent Administration of Parathyroid Hormone 1–34 Enhances Osteogenesis of Human Mesenchymal Stem Cells by Regulating Protein Kinase Cδ
Abstract
:1. Introduction
2. Results
2.1. Intermittent PTH (1–34) Treatment Enhances Osteogenesis
2.2. Intermittent PTH (1–34) Treatment Increases Early-Stage Osteoblast Activity during Osteogenic Differentiation
2.3. Intermittent PTH (1–34) Activates the PKC Pathway
2.4. PKC Is Involved in Intermittent PTH (1–34)-Enhanced Osteogenesis
2.5. PKCδ Mediates Intermittent PTH (1–34)-Enhanced Osteogenesis of hMSCs
3. Discussion
4. Materials and Methods
4.1. Cell Culture and Osteogenic Differentiation of hMSCs
4.2. PTH (1–34) Treatment
4.3. Inhibitors
4.4. Reverse Transcription and Quantitative Real-Time Polymerase Chain Reaction (RT-qPCR)
4.5. Alkaline Phosphatase (ALP) Staining
4.6. Alkaline Phosphatase (ALP) Activity Assay
4.7. Protein Kinase C (PKC) Kinase Activity Assay
4.8. Protein Kinase A (PKA) Kinase Activity Assay
4.9. Statistical Analysis
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
MSCs | Mesenchymal stromal cells |
PTH | Parathyroid hormone |
PKC | Protein kinase C |
PKCδ | Protein kinase C δ |
PKCα | Protein kinase C α |
RUNX2 | Runt-related transcription factor 2 |
COL1a1 | Collagen type 1 a1 |
ALP | Alkaline phosphatase |
OIM | Osteogenic induction medium |
ATF4 | Activating transcription factor 4 |
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Gene Name | Oligonucleotide Sequence | Probe Number |
---|---|---|
RUNX2 | 5′–CTACCACCCCGCTGTCTTC–3′/5′–CAGAGGTGGCAGTGTCATCA–3′ | 29 |
COL1a1 | 5′–ATGTTCAGCTTTGTGGACCTC–3′/5′–CTGTACGCAGGTGATTGGTG–3′ | 15 |
ALP | 5′–AGAACCCCAAAGGCTTCTTC–3′/5′–CTTGGCTTTTCCTTCATGGT–3′ | 31 |
Osterix | 5′–GACTGCAGAGCAGGTTCCTC-3′/5′–TAACCTGATGGGGTCATGGT–3′ | 43 |
PKCδ | 5′–TCGACTGGGAAAAACTGGAG–3′/5′–CTTGGTTGGTTCCCTTTCCA–3′ | 80 |
PKCα | 5’–ATTATCCCCGCTGGATCAC–3′/5′–CTCTGCTCCTTTGCCACAC–3′ | 83 |
ATF4 | 5′–TGGTCAGTCCCTCCAACAAC–3′/5′–CTATACCCAACAGGGCATCC–3′ | 88 |
GAPDH | 5′–GCTCTCTGCTCCTCCTGTTC–3′/5′–ACGACCAAATCCGTTGACTC–3′ | 60 |
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Kuo, S.-W.; Rimando, M.G.; Liu, Y.-S.; Lee, O.K. Intermittent Administration of Parathyroid Hormone 1–34 Enhances Osteogenesis of Human Mesenchymal Stem Cells by Regulating Protein Kinase Cδ. Int. J. Mol. Sci. 2017, 18, 2221. https://doi.org/10.3390/ijms18102221
Kuo S-W, Rimando MG, Liu Y-S, Lee OK. Intermittent Administration of Parathyroid Hormone 1–34 Enhances Osteogenesis of Human Mesenchymal Stem Cells by Regulating Protein Kinase Cδ. International Journal of Molecular Sciences. 2017; 18(10):2221. https://doi.org/10.3390/ijms18102221
Chicago/Turabian StyleKuo, Shu-Wen, Marilyn G. Rimando, Yi-Shiuan Liu, and Oscar K. Lee. 2017. "Intermittent Administration of Parathyroid Hormone 1–34 Enhances Osteogenesis of Human Mesenchymal Stem Cells by Regulating Protein Kinase Cδ" International Journal of Molecular Sciences 18, no. 10: 2221. https://doi.org/10.3390/ijms18102221