Potential Spermatogenesis Recovery with Bone Marrow Mesenchymal Stem Cells in an Azoospermic Rat Model
Abstract
:1. Introduction
2. Results
2.1. Cell Culture and Labeling
2.2. BMSCs Exhibit Multi-Lineage Differentiation Ability
2.3. Morpholgy Changes and Spermatogenenic Protein Expression of Induced BMSCs in Vitro
2.4. Recipient Rats
2.5. Donor Cells in Recipient Seminiferous Tubules
2.6. Expression of Spermatogenic Molecular Markers in Recipient Testicular Tissue
Group | Integrin-β1 | c-kit | GCNF | HSP90α |
---|---|---|---|---|
Nor | 0.77 ± 0.10 | 2.17 ± 0.50 | 1.84 ± 0.70 | 0.65 ± 0.17 |
0 day | 0.01 ± 0.01 * | 0.94 ± 0.19 * | 0.48 ± 0.13 * | 0.21 ± 0.08 * |
1 week | 0.11± 0.05 * | 0.62 ± 0.17 * | 0.27 ± 0.09 * | 0.45 ± 0.06 * |
2 weeks | 0.20 ± 0.08 * | 0.27 ± 0.08 * | 0.35 ± 0.09 * | 0.65 ± 0.13 |
4 weeks | 0.33 ± 0.12 * | 0.85 ± 0.14 * | 1.30 ± 0.23 | 0.76 ± 0.09 |
8 weeks | 0.69 ± 0.22 | 0.75 ± 0.24 * | 1.36 ± 0.25 | 0.71 ± 0.11 |
3. Discussion
4. Materials and Methods
4.1. Experimental Animals
4.2. BMSCs Collection, Culture and Differentiation Potential Test
4.3. Induction of BMSCs to Spermatogenic Differentiation in Vitro by Co-Culture with Testicular Sertoli Cells in Conditioned Media
4.4. Preparation of Donor MSCs for Transplantation in Vivo
4.5. Preparation of Recipient Rats: Busulfan-Induced Azoospermatism Model
4.6. BMSCs Transplantation and Testicular Tissue Collection
4.7. RNA Extraction and Quantitative Real-Time RT-PCR
4.8. SDS-PAGE and Western Blotting
Target Gene | Locus No. | Sequence of Primers | Product Size |
---|---|---|---|
Vasa | S75275 | F: 5'-GCGAGACTACATCTACAAC-3' | 135 bp |
R: 5'-GAGTATCTTCACAGTCATTA-3' | |||
SMAD1 | AF067727 | F: 5'-CTCATGTCATTTATTGCCG-3' | 138 bp |
R: 5'-CTCGCTTATAGTGGTAGGGA-3' | |||
Stella | BK001414 | F: 5'-CTATCATCGTCGTCAAAGG-3' | 177 bp |
R: 5'-CTCTGCTCAATCCGAACAA-3' | |||
Dazl | NM_001025742 | F: 5'-CGACGAAATCGGGAAGCTC-3' | 94 bp |
R: 5'-CACAACCTCACCATACTGGGAAA-3' | |||
GCNF | AJ783965 | F: 5'-CAACTGAACAAGCGGTATT-3' | 114 bp |
R: 5'-GATGTATCGGATCTCTGGC-3' | |||
c-kit | NM_022264 | F: 5'-TGCCCGAAACAAGTCATCTCC-3' | 112 bp |
R: 5'-GGCTGAGGGTTCAACTTTATCCA-3' | |||
β-actin | NM_031144 | F: 5'-GCTCGTCGTCGACAACGGCTC-3' | 353 bp |
R: 5'-CAAACATGATCTGGGTCATCTTCTC-3' |
4.9. Statistical Analysis
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Zhang, D.; Liu, X.; Peng, J.; He, D.; Lin, T.; Zhu, J.; Li, X.; Zhang, Y.; Wei, G. Potential Spermatogenesis Recovery with Bone Marrow Mesenchymal Stem Cells in an Azoospermic Rat Model. Int. J. Mol. Sci. 2014, 15, 13151-13165. https://doi.org/10.3390/ijms150813151
Zhang D, Liu X, Peng J, He D, Lin T, Zhu J, Li X, Zhang Y, Wei G. Potential Spermatogenesis Recovery with Bone Marrow Mesenchymal Stem Cells in an Azoospermic Rat Model. International Journal of Molecular Sciences. 2014; 15(8):13151-13165. https://doi.org/10.3390/ijms150813151
Chicago/Turabian StyleZhang, Deying, Xing Liu, Jinpu Peng, Dawei He, Tao Lin, Jing Zhu, Xuliang Li, Yuanyuan Zhang, and Guanghui Wei. 2014. "Potential Spermatogenesis Recovery with Bone Marrow Mesenchymal Stem Cells in an Azoospermic Rat Model" International Journal of Molecular Sciences 15, no. 8: 13151-13165. https://doi.org/10.3390/ijms150813151