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Differentiation of Human Spermatogonial Stem Cells Using a Human Decellularized Testicular Scaffold Supplemented by Platelet-Rich Plasma Publisher Pubmed



Salem M1 ; Feizollahi N1 ; Jabari A2 ; Golmohammadi MG3 ; Shirinsokhan A4 ; Ghanamigashti N5, 6 ; Bashghareh A1 ; Nikmahzar A1 ; Abbasi Y7 ; Naji M8 ; Abbasi M1
Authors
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Authors Affiliations
  1. 1. Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
  2. 2. Department of Obstetrics and Gynecology, Molud Infertility Center, Zahedan University of Medical Sciences, Zahedan, Iran
  3. 3. Department of Anatomy, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran
  4. 4. Department of Biology, Faculty of Sciences, Rasht Branch, Islamic Azad University, Rasht, Iran
  5. 5. Biomaterials Cluster, Bernal Institute, University of Limerick, Limerick, Ireland
  6. 6. School of Engineering, University of Limerick, Limerick, Ireland
  7. 7. School of Dentistry, Tehran University of Medical Sciences, Tehran, Iran
  8. 8. Urology and Nephrology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

Source: Artificial Organs Published:2023


Abstract

Background: Effective culture systems for attachment, migration, proliferation, and differentiation of spermatogonial stem cells (SSCs) can be a promising therapeutic modality for preserving male fertility. Decellularized extracellular matrix (ECM) from native testis tissue creates a local microenvironment for testicular cell culture. Furthermore, platelet-rich plasma (PRP) contains various growth factors for the proliferation and differentiation of SSCs. Methods: In this study, human testicular cells were isolated and cultured for 4 weeks, and SSCs were characterized using immunocytochemistry (ICC) and flow cytometry. Human testicular tissue was decellularized (0.3% SDS, 1% Triton), and the efficiency of the decellularization process was confirmed by histological staining and DNA content analysis. SSCs were cultured on the human decellularized testicular matrix (DTM) for 4 weeks. The viability and the expression of differentiation genes were evaluated by MTT and real-time polymerase chain reaction (PCR), respectively. Results: Histological evaluation and DNA content analysis showed that the components of ECM were preserved during decellularization. Our results showed that after 4 weeks of culture, the expression levels of BAX, BCL-2, PLZF, and SCP3 were unchanged, while the expression of PRM2 significantly increased in the cells cultured on DTM supplemented with PRP (ECM-PRP). In addition, the expression of GFRA1 was significantly decreased in the ECM group compared to the control and PRP groups. Furthermore, the MTT test indicated that viability was significantly enhanced in cells plated on DTM supplemented with PRP. Conclusion: Our study demonstrated that DTM supplemented with PRP can provide an effective culture system for the differentiation and viability of SSCs. © 2023 International Center for Artificial Organ and Transplantation (ICAOT) and Wiley Periodicals LLC.
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