IL-1b in the Secretomes of MSCs Seeded on Human Decellularized Allogeneic Bone Promotes Angiogenesis
Abstract
:1. Introduction
2. Results
2.1. MSC-CMs Promote Cell Growth
2.2. The Effect of MSC-CMs on Cell Migration
2.3. Analysis of Angiogenic Potential of MSC Secretomes by Tube Formation Assay
2.4. qPCR Profiling of Angiogenic Genes
2.5. Validation of Analyzed Results by IL-1b Neutralization
3. Discussion
4. Materials and Methods
4.1. MSC Culture
4.2. Processing of Allogeneic Bone
4.3. HUVEC Culture
4.4. Preparation of Conditioned Media
4.5. MTT Viability Assay
4.6. Wound Healing Assay
4.7. Tube Formation Assay
4.8. Real-Time Quantitative Polymerase Chain Reaction Analysis
4.9. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gene Name | Forward Primer (5′ to 3′) | Reverse Primer (5′ to 3′) | Accession No. | Product Size (bp) |
---|---|---|---|---|
ANG | ACTGGAACCCATCTCCAGGAACA | CAACACAGGCTCCCAGGAGGAA | NM_001097577 | 100 |
ANGPT1 | TCAGTGGCTGCAAAAACTTGAGA | CAGCATGGTAGCCGTGTGGT | NM_001146 | 103 |
ANGPT2 | GCTAAGGACCCCACTGTTGCTA | TCCATGTCACAGTAGGCCTTGAT | NM_001147 | 146 |
ARTN | GCTGTTTGAGCTTCGGGGGA | TGTTCCCCCACCCTCCTGTT | NM_057091 | 104 |
CCL13 | ACAGCAGCTTTCAACCCCCA | TGAAGCAGCAAGTAGATGGGACG | NM_005408 | 70 |
CCL3 | GGTGTCATCTTCCTAACCAAGCGAA | CTCAGGCACTCAGCTCCAGGTC | NM_002983 | 100 |
CCL7 | AGGCTGGAGAGCTACAGAAGGAC | CTGTGTGGGGTCAGCACAGAT | NM_006273 | 102 |
COL18A1 | CGACTTCCAGCCGGTGCTC | GACAGGAAGGCGCGGAAGG | NM_001379500 | 141 |
CSF2 | CGGAAACTTCCTGTGCAACCC | CCTCATCTGGCCGGTCTCACT | NM_000758 | 126 |
CSF3 | CTGGACAGTGCAGGAAGCCAC | TAGGTGGCACACAGCTTCTCCT | NM_172219 | 135 |
EDN1 | GCCAAAAAGACAAGAAGTGCTGG | TCCATAATGTCTTCAGCCCTGAGTT | NM_001955 | 70 |
EGF | AGGAGAACATCTCTCAACCACGAG | AGGCCTTGGAGGGAAGAACTTT | NM_001963 | 112 |
ENG | AGCCCCACAAGTCTTGCAGAA | CACGCAGCCCTTCGAGACCT | NM_001114753 | 105 |
FGF2 | CGGGTGCAGTGGCTCATGCCTATA | CGGGGTTTCACCAGGTTGGTCTTG | NM_002006 | 100 |
FGFR1 | GCATGGTGGGGTCGGTCATC | CTGGAGTCAGCAGACACTGTTACC | NM_023110 | 136 |
FLT1 | TGACCCACATTGGCCACCATC | GTGTAGTGCTGCATCCTTGTTGA | NM_002019 | 167 |
HGF | CAGCATCATCGAGGGAAGGTGACTC | CCCCTCACATGGTCCTGATCCAA | NM_000601 | 84 |
HIF1A | GACATCGCGGGGACCGATTC | CGCCGAGATCTGGCTGCAT | NM_001530 | 112 |
IGF1 | AGCTGGTGGATGCTCTTCAGTTCG | CACTCATCCACGATGCCTGTCTGA | NM_001111283 | 115 |
IGFBP1 | CAACCTCTGCACGCCCTCAC | TCTCCGTGCTCTCTGGGCTT | NM_000596 | 112 |
IGFBP2 | CTCCCTGCACATCCCCAACTG | CTTCCCGGTGTTGGGGTTCA | NM_000597 | 115 |
IGFBP3 | ACCACCAAGGGGAAGGAGGA | AGCTGCTGGTCATGTCCTTGG | NM_000598 | 130 |
IL-1A | TGTGACTGCCCAAGATGAAGACCA | TGCCGTGAGTTTCCCAGAAGAAGAG | NM_000575 | 113 |
IL-1B | GAAGTACCTGAGCTCGCCAGT | GCCTGAAGCCCTTGCTGTAGT | NM_000576 | 181 |
IL-4 | GAGAAGGACACTCGCTGCCTG | GAGGTTCCTGTCGAGCCGTT | NM_000589 | 93 |
IL-8 | GACCACACTGCGCCAACACAGAA | CCACAACCCTCTGCACCCAGTTT | NM_000584 | 96 |
KDR | CAAGTGGCTAAGGGCATGGA | ATTTCAAAGGGAGGCGAGCA | NM_002253 | 181 |
LEP | TAGGAATCGCAGCGCCAGC | TGTGTGAAATGTCATTGATCCTGGT | NM_000230 | 198 |
MMP8 | CCAGCAACTACTCACTCCCTCAAG | CAGGGTTTGGGTGTGCTTGGT | NM_002424 | 106 |
PDGFA | ATCGGGAAGAGGACACGGGAA | ATCTGGTTGGCTGCTTTAGGTGG | NM_002607 | 89 |
PDGFB | ACCACCTGGCATGCAAGTGTGAGAC | TCCGAATGGTCACCCGAGTTTGG | NM_002608 | 114 |
PDGFC | CTGAACCAGGGTTCTGCATCCACT | AGGGGGTAGCACTGAAGGACTCACA | NM_016205 | 80 |
PDGFD | GGAAGATTTCCAACCCGCAGCA | GTCCAGAGCATCCGCAATCAGAGT | NM_025208 | 124 |
PLAU | GGACCCCTCGTCTGTTCCCT | GGTGTGACTGCGGATCCAGG | NM_002658 | 138 |
PLG | GCTGACCGGACCGAATGTTT | GAGTTCGGTGGATTGGACTCTTCC | NM_000301 | 153 |
PRL | GGCTTCTAGAGGGCATGGAGC | CGTAGGCAGTGGAGCAGGTT | NM_000948 | 154 |
PTX3 | GGCAGACGCGAGCCGA | AAGCCTCATTGGTCTCACTGGATG | NM_002852 | 143 |
SERPINB5 | TGGAGGCCACGTTCTGTATGG | GGGTAGTAGGATGAACATGCTGAG | NM_002639 | 101 |
SERPINE1 | GCCCATGATGGCTCAGACCAA | CTGAGGGTGTCCCCGTGGTA | NM_000602 | 105 |
SERPINF1 | AGGCGAAGTCACCAAGTCCC | TCAAAGCCAGCCCGGTGTTC | NM_002615 | 123 |
TEK | TGCCACCATCACTCAGTATCAGCTC | TCCGCTGGTTGCTTGAGATTCTG | NM_000459 | 147 |
THBS1 | TGCTGGTGGTAGACTAGGGTTGTT | ATCCTGGGGGTTTTCTCAAGCC | NM_003246 | 173 |
THBS2 | GGGCGGCTGGGTCTATTTGT | GCACAGGGCATTGCCGGA | NM_003247 | 113 |
THPO | TGGGTCCTGGAGCCCTTCTC | GGAGGCGGCTTAGGCTCTTG | NM_000460 | 101 |
TIE1 | GTACGAGCTGATGCGTCAGTGCT | GCCCGCGTAAGTGAAGTTCTCA | NM_005424 | 145 |
VEGFA | CCATGCCAAGTGGTCCCAGG | GATGGCAGTAGCTGCGCTGA | NM_001025366 | 104 |
VWF | CAATGAGTTCCAACTGCAGCTCAGC | TGCCATCCCTCAGCATGAAGTCA | NM_000552 | 110 |
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Chen, Y.-C.; Fu, Y.-S.; Tsai, S.-W.; Wu, P.-K.; Chen, C.-M.; Chen, W.-M.; Chen, C.-F. IL-1b in the Secretomes of MSCs Seeded on Human Decellularized Allogeneic Bone Promotes Angiogenesis. Int. J. Mol. Sci. 2022, 23, 15301. https://doi.org/10.3390/ijms232315301
Chen Y-C, Fu Y-S, Tsai S-W, Wu P-K, Chen C-M, Chen W-M, Chen C-F. IL-1b in the Secretomes of MSCs Seeded on Human Decellularized Allogeneic Bone Promotes Angiogenesis. International Journal of Molecular Sciences. 2022; 23(23):15301. https://doi.org/10.3390/ijms232315301
Chicago/Turabian StyleChen, Yi-Chun, Yu-Show Fu, Shang-Wen Tsai, Po-Kuei Wu, Chao-Ming Chen, Wei-Ming Chen, and Cheng-Fong Chen. 2022. "IL-1b in the Secretomes of MSCs Seeded on Human Decellularized Allogeneic Bone Promotes Angiogenesis" International Journal of Molecular Sciences 23, no. 23: 15301. https://doi.org/10.3390/ijms232315301