Differentiation of human spermatogonial stem cells using a human decellularized testicular scaffold supplemented by platelet‐rich plasma

Author:

Salem Maryam1,Feizollahi Narjes1,Jabari Ayob2,Golmohammadi Mohammad Ghasem3,Shirinsokhan Armaghan4,Ghanami Gashti Nasrin56,Bashghareh Alieh1,Nikmahzar Aghbibi1,Abbasi Yasaman7,Naji Mohammad8ORCID,Abbasi Mehdi1

Affiliation:

1. Department of Anatomy, School of Medicine Tehran University of Medical Sciences Tehran Iran

2. Department of Obstetrics and Gynecology, Molud Infertility Center Zahedan University of Medical Sciences Zahedan Iran

3. Department of Anatomy, School of Medicine Ardabil University of Medical Sciences Ardabil Iran

4. Department of Biology, Faculty of Sciences, Rasht Branch Islamic Azad University Rasht Iran

5. Biomaterials Cluster, Bernal Institute University of Limerick, Limerick, Ireland Limerick Ireland

6. School of Engineering University of Limerick, Limerick, Ireland Limerick Ireland

7. School of Dentistry Tehran University of Medical Sciences Tehran Iran

8. Urology and Nephrology Research Center Shahid Beheshti University of Medical Sciences Tehran Iran

Abstract

AbstractBackgroundEffective 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.MethodsIn 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.ResultsHistological 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.ConclusionOur study demonstrated that DTM supplemented with PRP can provide an effective culture system for the differentiation and viability of SSCs.

Funder

Tehran University of Medical Sciences and Health Services

Publisher

Wiley

Subject

Biomedical Engineering,General Medicine,Biomaterials,Medicine (miscellaneous),Bioengineering

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