مقالات پذیرفته شده در نهمین کنگره بین المللی زیست پزشکی
Molecular Characterization and Gene Expression Analysis of Spermatogonial Stem Cells in Moghani Sheep
Molecular Characterization and Gene Expression Analysis of Spermatogonial Stem Cells in Moghani Sheep
Mostafa Ashrafi Osalou,1,*Hooman Ravaei,2
1. Department of Anatomical Science, Faculty of Medicine, Islamic Azad University, Ardabil Branch, Iran. 2. Young Researchers and Elite Club, Ardabil Branch, Islamic Azad University, Ardabil, Iran. Ardabil, Iran.
Introduction: Most investigations on spermatogonial stem cells (SSCs) have been restricted to laboratory animals, whereas sheep remain underexplored despite their dual significance in livestock productivity and their suitability as translational models in reproductive biotechnology. Given the growing prevalence of male infertility, particularly non-obstructive azoospermia, sheep SSCs offer a valuable platform for developing stem cell–based therapeutic strategies. This study aimed to isolate, characterize, and assess the gene expression profile of SSCs from the Moghani sheep breed, with a focus on their potential application in human reproductive medicine.
Methods: Testicular tissues were collected from Moghani sheep and processed through a four-step protocol: tissue collection and preparation, enzymatic digestion, filtration through cell strainers, and SSC seeding. Six SSC-specific markers (THY1, PLZF, PGP9.5, DAZL, C-KIT2, and VASA) were targeted by designing gene-specific primers. Following cell culture, cytoplasmic RNA was extracted, cDNA was synthesized via RT-PCR, amplified over 35 cycles, and analyzed through electrophoresis under ultraviolet light.
Results: Clear electrophoresis bands confirmed the expression of all six spermatogonial markers, validating the successful isolation and culture of SSCs from Moghani sheep. The cultured cells retained stemness, exhibited stable gene expression, and demonstrated long-term proliferation without signs of differentiation in vitro. These findings corroborate recent advances showing that optimized culture conditions and cryopreservation strategies preserve SSC functionality in sheep, thus broadening their research utility.
Conclusion: This study provides evidence that SSCs derived from Moghani sheep testes can be reliably isolated and propagated in vitro while maintaining expression of key germline markers. The translational relevance of these findings lies in their potential application to human infertility research: by serving as a robust model for spermatogonial biology, sheep SSCs could inform future therapeutic approaches for azoospermia and support the development of reproductive technologies aimed at restoring spermatogenesis in infertile men.