Alteration of spermatogenesis following spermatogonial stem cells transplantation in testicular torsion-detorsion mice

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 β 1 + ) 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 postmeiotic 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 Capsule SSCs transplantation increase relative expression of pre-/postmeiotic genes and improve sperm parameters and testis structure in testicular torsion-detorsion mice.Summary sentence SSCs transplantation increase relative expression of pre-/post-meiotic genes and improve sperm parameters and testis structure in the ischemic condition. 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.

Silk nanofibrous electrospun scaffold enhances differentiation of embryonic stem like cells derived from testis in to mature neuron

“…Nowadays, cell therapy has been widely accepted as an alternative therapeutic strategy in translational medicine. 1,24,25 Pluripotent stem cells are considered as promising source for cell engraftment in neuroregenerative medicine in regard to their utility to neural differentiation. 10 But, ethical and technical limitations associated with embryonic stem cells limits their clinical application.…”

“…Before neural induction, 2 × 10 5 cells were seeded on silk scaffold and cultured for 24 h. Neural differentiation method of ES like cells, for example, 4−/4+ Retinoic Acid technique was used for neural induction of ES like cells. Cells were exposed to RA at the first step to induce neuroprogenitor cells then, were cultured in the neurobasal medium supplemented with 20 ng/mL basic Fibroblastic Factor, 10 ng/mL Epidermal Growth Factor, B27 and N2 for 8–12 days . The differentiated cells were characterized for neuron biomarkers with Real time‐PCR and immunocytochemistry techniques.…”

“…Cells were exposed to RA at the first step to induce neuroprogenitor cells then, were cultured in the neurobasal medium supplemented with 20 ng/mL basic Fibroblastic Factor, 10 ng/mL Epidermal Growth Factor, B27 and N2 for 8-12 days. 24 The differentiated cells were characterized for neuron biomarkers with Real time-PCR and immunocytochemistry techniques.…”

Current approaches for the treatment of male infertility with stem cell therapy

“…Hermann et al () showed that after inducing infertility with alkylating chemotherapy, autologous, and allogeneic SSC transplantations into the testes of adult and prepubertal macaques, restored spermatogenesis and caused functional sperm production. Azizollahi et al () also demonstrated that testicular ischemia happens following testicular torsion and could be treated by auto‐transplantation of SSCs in mice through the up‐regulation of pre‐ and post‐meiotic genes expression, resulting in testicular function, and tissue improvement. Ogawa, Dobrinski, Avarbock, and Brinster () explained that the transplantation of SSCs from infertile steel ( Sl ) mice into infertile dominant white spotting ( W ) mice may generate spermatogenesis in the recipient mice.…”

“…potent adult germline stem cells (maGSC), is similar to ESCs and after meiosis they can form haploid male germ cells in vitro(Nolte et al, 2010) Hermann et al (2012). showed that after inducing infertility with alkylating chemotherapy, autologous, and allogeneic SSC transplantations into the testes of adult and prepubertal macaques, restored spermatogenesis and caused functional sperm production Azizollahi et al (2016). also demonstrated that testicular ischemia happens following testicular torsion and could be treated by auto-transplantation of SSCs in mice through the up-regulation of pre-and post-meiotic genes expression, resulting in testicular function, and tissue improvement Ogawa, Dobrinski, Avarbock, and Brinster (2000).…”