Saeid Azizollahi1, Reza Aflatoonian2, Mohammad Ali Sadighi Gilani3,4, Babak Behnam5,6,7,8, Nader Tajik9, Mohammad Asghari-Jafarabadi10, Hamid Reza Asgari1, Morteza Koruji11,12. 1. Department of Anatomical Sciences, School of Medicine, Iran University of Medical Sciences, Tehran, Iran. 2. Department of Endocrinology and Female Infertility, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran. 3. Department of Urology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran. 4. Department of Andrology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran. 5. Department of Medical Genetics and Molecular Biology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran. 6. NIH Undiagnosed Diseases Program, NIH, Office of the Director, Bethesda, MD, 20892, USA. 7. National Human Genome Research Institute, NIH, Bethesda, MD, 20892, USA. 8. Cellular and Molecular Research Center, Iran University of Medical Sciences (IUMS), Hemmat Highway, P. O. Box 14155-5983, Tehran, Iran. 9. Department of Immunology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran. 10. Road Traffic Injury Research Center, Tabriz University of Medical Sciences, Tabriz, Iran. 11. Department of Anatomical Sciences, School of Medicine, Iran University of Medical Sciences, Tehran, Iran. koruji.m@iums.ac.ir. 12. Cellular and Molecular Research Center, Iran University of Medical Sciences (IUMS), Hemmat Highway, P. O. Box 14155-5983, Tehran, Iran. koruji.m@iums.ac.ir.
Abstract
PURPOSE: Testicular ischemia is the main consequence of testicular torsion, in both clinical and experimental aspects. Preservation and auto-transplantation of spermatogonial stem cells (SSCs) could be a new treatment for infertility in testicular ischemia following testicular torsion. METHODS: To apply the idea in this study, animals were randomly divided into four groups of control, sham, with torsion, and with torsion followed by transplantation (TT). Isolated SSCs from neonatal mice were cultured and identified by flow cytometry (C-KIT(-), INTEGRIN β1 (+)) and RT-PCR (Reverse transcription polymerase chain reaction) for specific spermatogonial cell markers (Oct4, Gfrα-1, Plzf, Vasa, Itgα 6 , and Itgβ 1 ). SSCs were transplanted upon a 2-h testicular torsion in the TT group. Cultured cells were transplanted into ischemia reperfusion testicle 2 weeks post-testicular torsion. Eight weeks after SSCs transplantation, the SSCs-transplanted testes and epididymis were removed for sperm analysis, weight and histopathological evaluation, and pre- and post-meiotic gene expression assessment by qRT-PCR. RESULTS: Our findings indicated that all evaluated parameters (epididymal sperm profile, Johnsen score, Plzf, Gfrα-1, Scp-1, Tekt-1 expressions, and histopathological profile) were significantly decreased following testicular torsion (group 3) when compared to the control group (p ≤ 0.05). However, all abovementioned parameters showed a significant increase/improvement in torsion-transplantation group compared to torsion group. However, these parameters in the TT group were significantly lower in the sham and control groups (p ≤ 0.05). CONCLUSION: SSCs transplantation could up-regulate the expression of pre- and post-meiotic genes in testicular ischemia, which resulted in improvement of both testicular function and structure after testicular torsion.
PURPOSE:Testicular ischemia is the main consequence of testicular torsion, in both clinical and experimental aspects. Preservation and auto-transplantation of spermatogonial stem cells (SSCs) could be a new treatment for infertility in testicular ischemia following testicular torsion. METHODS: To apply the idea in this study, animals were randomly divided into four groups of control, sham, with torsion, and with torsion followed by transplantation (TT). Isolated SSCs from neonatal mice were cultured and identified by flow cytometry (C-KIT(-), INTEGRIN β1 (+)) and RT-PCR (Reverse transcription polymerase chain reaction) for specific spermatogonial cell markers (Oct4, Gfrα-1, Plzf, Vasa, Itgα 6 , and Itgβ 1 ). SSCs were transplanted upon a 2-h testicular torsion in the TT group. Cultured cells were transplanted into ischemia reperfusion testicle 2 weeks post-testicular torsion. Eight weeks after SSCs transplantation, the SSCs-transplanted testes and epididymis were removed for sperm analysis, weight and histopathological evaluation, and pre- and post-meiotic gene expression assessment by qRT-PCR. RESULTS: Our findings indicated that all evaluated parameters (epididymal sperm profile, Johnsen score, Plzf, Gfrα-1, Scp-1, Tekt-1 expressions, and histopathological profile) were significantly decreased following testicular torsion (group 3) when compared to the control group (p ≤ 0.05). However, all abovementioned parameters showed a significant increase/improvement in torsion-transplantation group compared to torsion group. However, these parameters in the TT group were significantly lower in the sham and control groups (p ≤ 0.05). CONCLUSION: SSCs transplantation could up-regulate the expression of pre- and post-meiotic genes in testicular ischemia, which resulted in improvement of both testicular function and structure after testicular torsion.
Entities:
Keywords:
Pre- and post-meiotic genes; SSCs transplantation; Testicular torsion
Authors: F Izadyar; K Den Ouden; T A E Stout; J Stout; J Coret; D P K Lankveld; T J P Spoormakers; B Colenbrander; J K Oldenbroek; K D Van der Ploeg; H Woelders; H B Kal; D G De Rooij Journal: Reproduction Date: 2003-12 Impact factor: 3.906
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