Biological Characteristics and Osteogenic Differentiation of Ovine Bone Marrow Derived Mesenchymal Stem Cells Stimulated with FGF-2 and BMP-2
Abstract
:1. Introduction
2. Results
2.1. Morphology of Sheep BM-MSCs
2.2. Cell Proliferation and Doubling Time Analyzed with the MTT Assay
2.3. Immunophenotype of Sheep BM-MSCs in Different Culture Conditions
2.4. CD90 Expression Level in Sheep BM-MSCs
2.5. Assessment of Osteogenic Differentiation Markers
2.6. Effect of BMP-2 and FGF-2 on mRNA Expression in Genes Involved in Osteogenic Differentiation
2.7. Differentiation Potential of Sheep BM-MSCs
2.8. Secretion Profile of Sheep BM-MSCs
3. Discussion
4. Materials and Methods
4.1. BM-MSC Isolation and Culture
4.2. BMP-2 and FGF-2 Supplementation
4.3. Cell Proliferation and Doubling Time Calculation
4.4. Flow Cytometry
4.5. Immunofluorescence Staining
4.6. RT-PCR (Reverse Transcription Polymerase Chain Reaction)
4.7. Real-Time qRT-PCR
4.8. Multilineage Differentiation
4.9. Sheep Cytokine Array
4.10. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
AIF | allograft inflammatory factor |
BM-MSCs | bone marrow derived mesenchymal stem cells |
BMP-2 | bone morphogenetic protein 2 |
ColI | collagen type I |
DMSO | dimethyl sulfoxide |
FGF-2 | fibroblast growth factor 2 |
HLA DR | Human Leukocyte Antigen–DR isotype |
IL-8 | interleukin-8 |
MIG | monokine induced by gamma interferon |
MSCs | mesenchymal stem cells |
Ocl | osteocalcin |
Opn | osteopontin |
Osx | osterix |
RANTES | Regulated on Activation, Normal T-cell Expressed and Secreted |
Runx2 | Runt related transcription factor 2 |
sFRP-3 | secreted frizzled-related protein 3 |
TNF-α | tumor necrosis factor α |
VEGF-A | vascular endothelial growth factor A |
αMEM | Minimum Essential Medium α-transformation |
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Gromolak, S.; Krawczenko, A.; Antończyk, A.; Buczak, K.; Kiełbowicz, Z.; Klimczak, A. Biological Characteristics and Osteogenic Differentiation of Ovine Bone Marrow Derived Mesenchymal Stem Cells Stimulated with FGF-2 and BMP-2. Int. J. Mol. Sci. 2020, 21, 9726. https://doi.org/10.3390/ijms21249726
Gromolak S, Krawczenko A, Antończyk A, Buczak K, Kiełbowicz Z, Klimczak A. Biological Characteristics and Osteogenic Differentiation of Ovine Bone Marrow Derived Mesenchymal Stem Cells Stimulated with FGF-2 and BMP-2. International Journal of Molecular Sciences. 2020; 21(24):9726. https://doi.org/10.3390/ijms21249726
Chicago/Turabian StyleGromolak, Sandra, Agnieszka Krawczenko, Agnieszka Antończyk, Krzysztof Buczak, Zdzisław Kiełbowicz, and Aleksandra Klimczak. 2020. "Biological Characteristics and Osteogenic Differentiation of Ovine Bone Marrow Derived Mesenchymal Stem Cells Stimulated with FGF-2 and BMP-2" International Journal of Molecular Sciences 21, no. 24: 9726. https://doi.org/10.3390/ijms21249726