Volume 29, Issue 3 (September 2025)
Physiol Pharmacol 2025, 29(3): 293-305 |
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Soltani R, Abbaszadeh H, Norouzian M, Aghajanpour F, Afshar A, Aliaghaei A, et al . Sertoli cell-conditioned medium restores spermatogenesis in mice induced by scrotal hyperthermia. Physiol Pharmacol 2025; 29 (3) : 7
URL: http://ppj.phypha.ir/article-1-2281-en.html
URL: http://ppj.phypha.ir/article-1-2281-en.html
Reza Soltani 
, Hojjat-Allah Abbaszadeh , Mohsen Norouzian , Fakhroddin Aghajanpour , Azar Afshar , Abbas Aliaghaei , Nafiseh Moeinian , Ali Dehghani nejad , Fatemeh Fadaei Fathabadi 
, Mohammad-Amin Abdollahifar
, Hojjat-Allah Abbaszadeh , Mohsen Norouzian , Fakhroddin Aghajanpour , Azar Afshar , Abbas Aliaghaei , Nafiseh Moeinian , Ali Dehghani nejad , Fatemeh Fadaei Fathabadi , Mohammad-Amin Abdollahifar
Abstract: (2522 Views)
Introduction: Spermatogenesis is a complex physiological process susceptible to various influencing factors, among which scrotal hyperthermia is noteworthy. This investigation assesses the impact of conditioned medium derived from Sertoli cells on spermatogenic activity within the testicular tissue of adult male mice after exposure to scrotal hyperthermia.
Methods: A cohort of 40 adult male NMRI mice was employed in this research, and they were allocated ran-domly into four distinct groups as follows: (1) the control group comprising untreated animals, (2) the hyperthermia group subjected to scrotal hyperthermia through immersion in water at a tempera-ture of 43°C for 20 minutes, administered five times every other day, (3) the DMEM group receiv-ing 10μl of Dulbecco’s Modified Eagle Medium (DMEM), and (4) the Sertoli cell-conditioned me-dium (SCCM) group, administered 10μl of SCCM. After the induction of hyperthermia, intraperito-neal injections of SCCM were administered to the mice daily for 35 days. Samples of sperm were gathered from the epididymal tail, and the testis tissue was subsequently harvested for histological examination and molecular analysis.
Results: Our findings revealed that the administration of SCCM yielded significant increases in the quanti-ties of sperm and germ cells compared to the other groups. Additionally, the relative gene expres-sion levels of receptor tyrosine kinase (C-kit), stimulated by retinoic acid 8 (Stra8), and proliferat-ing cell nuclear antigen (Pcna) exhibited a substantial elevation within the SCCM group compared to the other experimental groups.
Conclusion: The utilization of SCCM holds promise as a therapeutic intervention for addressing infertility con-cerns, and it presents potential applications within the realms of the field of reproductive and re-generative medicine.
Article number: 7
Type of Manuscript: Experimental research article |
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References
1. Afshar A, Aliaghaei A, Nazarian H, Abbaszadeh H-A, Naserzadeh P, Fathabadi F F, et al. Curcumin-loaded iron particle improvement of spermatogenesis in azoospermic mouse induced by long-term scrotal hyperthermia. Reproductive Sciences 2021; 28: 371-380. [DOI:10.1007/s43032-020-00288-2]
2. Ahmad G, Moinard N, Esquerré-Lamare C, Mieusset R, Bujan L. Mild induced testicular and epididymal hyperthermia alters sperm chromatin integrity in men. Fertility and Sterility 2012; 97: 546-553. [DOI:10.1016/j.fertnstert.2011.12.025]
3. Aitken R J. Impact of oxidative stress on male and female germ cells: implications for fertility. Reproduction 2020; 159: R189-R201. [DOI:10.1530/REP-19-0452]
4. Anderson E L, Baltus A E, Roepers-Gajadien H L, Hassold T J, de Rooij D G, van Pelt A M, et al. Stra8 and its inducer, retinoic acid, regulate meiotic initiation in both spermatogenesis and oogenesis in mice. Proceedings of the National Academy of Sciences 2008; 105: 14976-14980. [DOI:10.1073/pnas.0807297105]
5. Cho C-L, Esteves S C, Agarwal A. Novel insights into the pathophysiology of varicocele and its association with reactive oxygen species and sperm DNA fragmentation. Asian Journal of Andrology 2016; 18: 186. [DOI:10.4103/1008-682X.170441]
6. Durairajanayagam D, Agarwal A, Ong C. Causes, effects and molecular mechanisms of testicular heat stress. Reproductive Biomedicine Online 2015; 30: 14-27. [DOI:10.1016/j.rbmo.2014.09.018]
7. Endo T, Freinkman E, de Rooij D G, Page D C. Periodic production of retinoic acid by meiotic and somatic cells coordinates four transitions in mouse spermatogenesis. Proceedings of the National Academy of Sciences 2017; 114: E10132-E10141. [DOI:10.1073/pnas.1710837114]
8. Endo T, Romer K A, Anderson E L, Baltus A E, de Rooij D G, Page D C. Periodic retinoic acid-STRA8 signaling intersects with periodic germ-cell competencies to regulate spermatogenesis. Proceedings of the National Academy of Sciences 2015; 112: E2347-E2356. [DOI:10.1073/pnas.1505683112]
9. Engel K M, Baumann S, Blaurock J, Rolle-Kampczyk U, Schiller J, von Bergen M, et al. Differences in the sperm metabolomes of smoking and nonsmoking men. Biology of Reproduction 2021; 105: 1484-1493. [DOI:10.1093/biolre/ioab179]
10. Feng C-W, Bowles J, Koopman P. Control of mammalian germ cell entry into meiosis. Molecular and Cellular Endocrinology 2014; 382: 488-497. [DOI:10.1016/j.mce.2013.09.026]
11. Fujioka H, Fujisawa M, Tatsumi N, Kanzaki M, Okuda Y, Okada H, et al. Sertoli cells inhibited apoptosis of pachytene spermatocytes and round spermatids. Endocrine Research 2001; 27: 75-90. [DOI:10.1081/ERC-100107171]
12. Geens M, Sermon K D, Van de Velde H, Tournaye H. Sertoli cell-conditioned medium induces germ cell differentiation in human embryonic stem cells. Journal of Assisted Reproduction And Genetics 2011; 28: 471-480. [DOI:10.1007/s10815-011-9541-9]
13. Ghaem Maghami R, Mirzapour T, Bayrami A. Differentiation of mesenchymal stem cells to germ-like cells under induction of Sertoli cell-conditioned medium and retinoic acid. Andrologia 2018; 50: e12887. [DOI:10.1111/and.12887]
14. Goshadezehn P, Babaei-Balderlou F, Razi M, Najafi G-R, Abtahi-Foroushani M. A caffeine pre-treatment and sole effect of bone-marrow mesenchymal stem cells-derived conditioned media on hyperglycemia-suppressed fertilization. Biomedicine & Pharmacotherapy 2023; 165: 115130. [DOI:10.1016/j.biopha.2023.115130]
15. Grootenhuis A, Timmerman M, Hordijk P, de Iong F. Inhibin in immature rat Sertoli cell conditioned medium: a 32 kDa αβ-B dimer. Molecular and Cellular Endocrinology 1990; 70: 109-116. [DOI:10.1016/0303-7207(90)90064-F]
16. Gundersen H, Bendtsen T F, Korbo L, Marcussen N, Møller A, Nielsen K, et al. Some new, simple and efficient stereological methods and their use in pathological research and diagnosis. Apmis 1988; 96: 379-394. [DOI:10.1111/j.1699-0463.1988.tb05320.x]
17. Hakovirta H, Yan W, Kaleva M, Zhang F, Vänttinen K, Morris P L, et al. Function of stem cell factor as a survival factor of spermatogonia and localization of messenger ribonucleic acid in the rat seminiferous epithelium. Endocrinology 1999; 140: 1492-1498. [DOI:10.1210/endo.140.3.6589]
18. Helsel A, Griswold M D. Retinoic acid signaling and the cycle of the seminiferous epithelium. Current Opinion in Endocrine and Metabolic Research 2019; 6: 1-6. [DOI:10.1016/j.coemr.2019.01.001]
19. Hengartner M O. The biochemistry of apoptosis. Nature 2000; 407: 770-776. [DOI:10.1038/35037710]
20. Hikim A P S, Lue Y, Yamamoto C M, Vera Y, Rodriguez S, Yen P H, et al. Key apoptotic pathways for heat-induced programmed germ cell death in the testis. Endocrinology 2003; 144: 3167-3175. [DOI:10.1210/en.2003-0175]
21. Izadpanah M, Alizadeh R, Minaee M B, Heydari L, Babatunde A, Abbasi M. The effects of curcumin on sperm parameters and nitric oxide production in varicocelized rats Efectos de la cúrcuma sobre los pará metros espermá ticos y producción de óxido nÃtrico en ratas varicocelizadas. International Journal of Morphology 2015; 33: 1530-1535. [DOI:10.4067/S0717-95022015000400055]
22. Kanatsu-Shinohara M, Inoue K, Takashima S, Takehashi M, Ogonuki N, Morimoto H, et al. Reconstitution of mouse spermatogonial stem cell niches in culture. Cell Stem Cell 2012; 11: 567-578. [DOI:10.1016/j.stem.2012.06.011]
23. Kazemeini S K, Emtiazy M, Owlia F, Khani P. Causes of infertility in view of Iranian traditional medicine: A review. International Journal of Reproductive Biomedicine 2017; 15: 187. [DOI:10.29252/ijrm.15.4.187]
24. Khanehzad M, Abbaszadeh R, Holakuyee M, Modarressi M H, Nourashrafeddin S M. FSH regulates RA signaling to commit spermatogonia into differentiation pathway and meiosis. Reproductive Biology and Endocrinology 2021; 19: 1-19. [DOI:10.1186/s12958-020-00686-w]
25. Li X, Zhu Q, Wen Z, Yuan K, Su Z, Wang Y, et al. Androgen and luteinizing hormone stimulate the function of rat immature leydig cells through different transcription signals. Frontiers in Endocrinology 2021; 12: 599149. [DOI:10.3389/fendo.2021.599149]
26. Maharlouei N, Morshed Behbahani B, Doryanizadeh L, Kazemi M. Prevalence and pattern of infertility in Iran: A systematic review and meta-analysis study. Women’s Health Bulletin 2021; 8: 63-71.
27. Maloy S, Hughes K. Brenner’s encyclopedia of genetics: Academic Press, 2013.
28. Meng J, Holdcraft R W, Shima J E, Griswold M D, Braun R E. Androgens regulate the permeability of the blood-testis barrier. Proceedings of the National Academy of Sciences 2005; 102: 16696-16700. [DOI:10.1073/pnas.0506084102]
29. Mieusset R, Bujan L, Massat G, Mansat A, Pontonnier F. Andrology: clinical and biological characteristics of infertile men with a history of cryptorchidism. Human Reproduction 1995; 10: 613-619. [DOI:10.1093/oxfordjournals.humrep.a135998]
30. Mínguez-Alarcón L, Gaskins A J, Chiu Y-H, Messerlian C, Williams P L, Ford J B, et al. Type of underwear worn and markers of testicular function among men attending a fertility center. Human Reproduction 2018; 33: 1749-1756. [DOI:10.1093/humrep/dey259]
31. Monageng E, Offor U, Takalani N B, Mohlala K, Opuwari C S. A Review on the Impact of Oxidative Stress and Medicinal Plants on Leydig Cells. Antioxidants 2023; 12: 1559. [DOI:10.3390/antiox12081559]
32. Monfared M H, Minaee B, Rastegar T, Khrazinejad E, Barbarestani M. Sertoli cell condition medium can induce germ like cells from bone marrow derived mesenchymal stem cells. Iranian Journal of bBasic Medical Sciences 2016; 19: 1186.
33. Nazifi S, Nazari H, Hassanpour H, Ahmadi E, Afzali A. Co-culturing or conditioned medium of Sertoli cells: Which one supports in vitro maturation of bovine oocytes and developmental competency of resulting embryos? Veterinary Medicine and Science 2022; 8: 2646-2654. [DOI:10.1002/vms3.939]
34. O’Donnell L, Smith L B, Rebourcet D. Sertoli cells as key drivers of testis function. Journal 2022; 121: 2-9. [DOI:10.1016/j.semcdb.2021.06.016]
35. Panahi S, Karamian A, Sajadi E, Aliaghaei A, Nazarian H, Abdi S, et al. Sertoli cell-conditioned medium restores spermatogenesis in azoospermic mouse testis. Cell and Tissue Research 2020; 379: 577-587. [DOI:10.1007/s00441-019-03092-w]
36. Paul C, Teng S, Saunders P T. A single, mild, transient scrotal heat stress causes hypoxia and oxidative stress in mouse testes, which induces germ cell death. Biology ofReproduction 2009; 80: 913-919. [DOI:10.1095/biolreprod.108.071779]
37. Pirani M, Novin M G, Abdollahifar M-A, Piryaei A, Kuroshli Z, Mofarahe Z S. Protective effects of fisetin in the mice induced by long-term scrotal hyperthermia. Reproductive Sciences 2021; 28: 3123-3136. [DOI:10.1007/s43032-021-00615-1]
38. Rebourcet D, Darbey A, Monteiro A, Soffientini U, Tsai Y T, Handel I, et al. Sertoli cell number defines and predicts germ and Leydig cell population sizes in the adult mouse testis. Endocrinology 2017; 158: 2955-2969. [DOI:10.1210/en.2017-00196]
39. Rockett J C, Mapp F L, Garges J B, Luft J C, Mori C, Dix D J. Effects of hyperthermia on spermatogenesis, apoptosis, gene expression, and fertility in adult male mice. Biology of Reproduction 2001; 65: 229-239. [DOI:10.1095/biolreprod65.1.229]
40. Rosa I, Marini M, Guasti D, Ibba-Manneschi L, Manetti M. Morphological evidence of telocytes in human synovium. Sciences Reports 2018; 8: 3581. [DOI:10.1038/s41598-018-22067-5]
41. Shaha C, Tripathi R, Mishra D P. Male germ cell apoptosis: regulation and biology. Philosophical Transactions of the Royal Society B: Biological Sciences 2010; 365: 1501-1515. [DOI:10.1098/rstb.2009.0124]
42. Sharma M, Leslie S W. Azoospermia. StatPearls 2022.
43. Shen X, Niu C, Guo J, Xia M, Xia J, Hu Y, et al. Stra8 may inhibit apoptosis during mouse spermatogenesis via the AKT signaling pathway. International Journal of Molecular Medicine 2018; 42: 2819-2830. [DOI:10.3892/ijmm.2018.3825]
44. Skinner M K. Sertoli cell secreted regulatory factors. Sertoli Cell Biology 2005; 1: 107-120. [DOI:10.1016/B978-012647751-1/50009-X]
45. Takashima S, Kanatsu-Shinohara M, Tanaka T, Morimoto H, Inoue K, Ogonuki N, et al. Functional differences between GDNF-dependent and FGF2-dependent mouse spermatogonial stem cell self-renewal. Stem Cell Reports 2015; 4: 489-502. [DOI:10.1016/j.stemcr.2015.01.010]
46. Tousson E, Ali E M, Ibrahim W, Mansour M A. Proliferating cell nuclear antigen as a molecular biomarker for spermatogenesis in PTU-induced hypothyroidism of rats. Reproductive Sciences 2011; 18: 679-686. [DOI:10.1177/1933719110395401]
47. Wang Z, Cai F, Chen X, Luo M, Hu L, Lu Y. The role of mitochondria-derived reactive oxygen species in hyperthermia-induced platelet apoptosis. PloS One 2013; 8: e75044. [DOI:10.1371/journal.pone.0075044]
48. Widlak W, Vydra N. The role of heat shock factors in mammalian spermatogenesis. Advances in Anatomy, Embryology and Cell Biology 2017: 45-65. [DOI:10.1007/978-3-319-51409-3_3]
49. Wu N, Murono E P. A Sertoli cell-secreted paracrine factor (s) stimulates proliferation and inhibits steroidogenesis of rat Leydig cells. Molecular and Cellular Endocrinology 1994; 106: 99-109. [DOI:10.1016/0303-7207(94)90191-0]
50. Yadav S K, Pandey A, Kumar L, Devi A, Kushwaha B, Vishvkarma R, et al. The thermo-sensitive gene expression signatures of spermatogenesis. Reproductive Biology and Endocrinology 2018; 16: 1-22. [DOI:10.1186/s12958-018-0372-8]
51. Yan W, Suominen J, Toppari J. Stem cell factor protects germ cells from apoptosis in vitro. Journal of Cell Science 2000; 113: 161-168. [DOI:10.1242/jcs.113.1.161]
52. Zhang L, Tang J, Haines C J, Feng H, Lai L, Teng X, et al. c-kit and its related genes in spermatogonial differentiation. Spermatogenesis 2011; 1: 186-194. [DOI:10.4161/spmg.1.3.17760]
53. Zhang L, Tang J, Haines C J, Feng H, Lai L, Teng X, et al. c-kit expression profile and regulatory factors during spermatogonial stem cell differentiation. BMC Developmental Biology 2013; 13: 1-14. [DOI:10.1186/1471-213X-13-38]
54. Ziaeipour S, Rezaei F, Piryaei A, Abdi S, Moradi A, Ghasemi A, et al. Hyperthermia versus busulfan: Finding the effective method in animal model of azoospermia induction. Andrologia 2019; 51: e13438. [DOI:10.1111/and.13438]
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