Characterization of Osteogenesis and Chondrogenesis of Human Decellularized Allogeneic Bone with Mesenchymal Stem Cells Derived from Bone Marrow, Adipose Tissue, and Wharton’s Jelly
Abstract
:1. Introduction
2. Results
2.1. Characterization of hMSCs and hDCB
2.2. Proliferation and Adhesion of hMSCs on hDCB Blocks
2.3. Production of Osteogenic Proteins and Calcium Deposition of hMSCs on hDCB Blocks
2.4. Production of Chondrogenic Proteins and Proteoglycans of hMSCs on hDCB Blocks
2.5. Comparison of Osteogenic and Chondrogenic Gene Expression Levels of hMSCs on hDCB Blocks Using Real-Time RT-qPCR
3. Discussion
4. Conclusions
5. Materials and Methods
5.1. Isolation and Expansion of Human MSCs
5.2. Processing of Allogeneic Bone
5.3. Characterization of hDCB Block
5.4. Osteogenic and Chondrogenic Differentiation of hMSCs
5.5. MTT Assay
5.6. Immunofluorescence
5.7. Alizarin Red S Staining
5.8. Safranin O Staining
5.9. Real-Time Quantitative Polymerase Chain Reaction Analysis of Osteogenic and Chondrogenic Genes
5.10. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Target Gene | Directions | Sequences | Accession No. | Product Size |
---|---|---|---|---|
ALPL | Forward | ACAGATGCCAACTTCCCACACG | NM_001200 | 112 bp |
Reverse | GCGGCAGACTTTGGTTTCTTGG | |||
COL1A1 | Forward | TCCCCTCCACTCCTTCCCAAA | NM_000088 | 146 bp |
Reverse | GGCCACTTGGGTGTTTGAGCA | |||
COL2A1 | Forward | TGGCTGACCTGACCTGATGTCC | NM_001844 | 95 bp |
Reverse | TGCAGTCTGCCCAGTTCAGGTC | |||
COL10A1 | Forward | AGGCCCACTACCCAACACCAAGA | NM_000493 | 161 bp |
Reverse | CGTAGCCTGGTTTTCCTGGTGGTC | |||
GAPDH | Forward | TGAGCACCAGGTGGTCTCCTCTGAC | NM_001256799 | 147 bp |
Reverse | TCCACCACCCTGTTGCTGTAGCCA | |||
Reverse | TGGGAGCAGCTGGGATGATG | |||
RUNX2 | Forward | ATGACGTCCCCGTCCATCCA | NM_001024630 | 135 bp |
Reverse | GGAAGGCCAGAGGCAGAAGTCA | |||
SOX9 | Forward | CCAAGCGCATTACCCACTTGTG | NM_000346 | 130 bp |
Reverse | CGATTCTCCATCATCCTCCACG |
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Chen, C.-F.; Chen, Y.-C.; Fu, Y.-S.; Tsai, S.-W.; Wu, P.-K.; Chen, C.-M.; Chang, M.-C.; Chen, W.-M. Characterization of Osteogenesis and Chondrogenesis of Human Decellularized Allogeneic Bone with Mesenchymal Stem Cells Derived from Bone Marrow, Adipose Tissue, and Wharton’s Jelly. Int. J. Mol. Sci. 2021, 22, 8987. https://doi.org/10.3390/ijms22168987
Chen C-F, Chen Y-C, Fu Y-S, Tsai S-W, Wu P-K, Chen C-M, Chang M-C, Chen W-M. Characterization of Osteogenesis and Chondrogenesis of Human Decellularized Allogeneic Bone with Mesenchymal Stem Cells Derived from Bone Marrow, Adipose Tissue, and Wharton’s Jelly. International Journal of Molecular Sciences. 2021; 22(16):8987. https://doi.org/10.3390/ijms22168987
Chicago/Turabian StyleChen, Cheng-Fong, Yi-Chun Chen, Yu-Show Fu, Shang-Wen Tsai, Po-Kuei Wu, Chao-Ming Chen, Ming-Chau Chang, and Wei-Ming Chen. 2021. "Characterization of Osteogenesis and Chondrogenesis of Human Decellularized Allogeneic Bone with Mesenchymal Stem Cells Derived from Bone Marrow, Adipose Tissue, and Wharton’s Jelly" International Journal of Molecular Sciences 22, no. 16: 8987. https://doi.org/10.3390/ijms22168987