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Sertoli cell–conditioned medium restores spermatogenesis in azoospermic mouse testis

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The current study evaluates potential applications of Sertoli cell (SC)–conditioned medium (CM) and explores the effects of the conditioned medium on the spermatogenesis process in azoospermic mice. For this study, 40 adult mice (28–30 g) were divided into 4 experimental groups: (1) control, (2) DMSO 2% (10 μl), (3) busulfan (40 mg/kg single dose) and (4) busulfan/CM (10 μl). SCs were isolated from 4-week-old mouse testes. After using anesthetics, 10 μl of CM was injected over 3–5 min into each testis and subsequently, sperm samples were collected from the tail of the epididymis. Afterward, the animals were euthanized and testis samples were taken for histopathology experiments and RNA extraction in order to examine the expression of c-kit, STRA8 and PCNA genes. The data showed that CM notably increased the total sperm count and the number of testicular cells, such as spermatogonia, primary spermatocytes, round spermatids, SCs and Leydig cells compared with the control, DMSO and busulfan groups. Furthermore, the results showed that expression of c-kit and STRA8 was significantly decreased in the busulfan and busulfan/SC groups at 8 weeks after the last injection (p < 0.001) but no significant difference was found for PCNA compared with the control and DMSO groups (p < 0.05). These findings suggest that the Sertoli cell–conditioned medium may be beneficial as a practical approach for therapeutic strategies in reproductive and regenerative medicine.

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This article has been extracted from the thesis by Mrs. Sakineh Panahi.


This study was financially supported by the Research Vice Chancellor of Shahid Beheshti University of Medical Sciences, Tehran, Iran (Registration No. 1395.115).

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Authors and Affiliations



MAA and SP designed this study and conducted the stereological study and provided the clinical data and sample. AK and ES carried out the animal model and immunohistochemistry. AA and HN performed the statistical analysis. SA and SD wrote and drafted the manuscript. SP carried out the real-time PCR. VE is responsible for the English language and grammar editing. RMF helped in writing the draft of the manuscript and helped in designing this study. All authors read and approved the final manuscript.

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Correspondence to Mohammad-Amin Abdollahifar or Reza Mastery Farahani.

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The authors declare that they have no conflict of interest.

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The experimental protocol and all procedures performed in studies involving the use of animals were reviewed and approved by the Ethical Committee at Shahid Beheshti University of Medical Sciences (IR. SBMU. SM.REC.1396.184).

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Panahi, S., Karamian, A., Sajadi, E. et al. Sertoli cell–conditioned medium restores spermatogenesis in azoospermic mouse testis. Cell Tissue Res 379, 577–587 (2020).

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