<i>In vitro</i>culture of testicular germ cells: Regulatory factors and limitations

Huleihel, Mahmoud; Abuelhija, Mahmoud; Lunenfeld, Eitan

doi:10.1080/08977190701783400

Cited by 50 publications

(61 citation statements)

References 157 publications

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“…Interestingly, coculture of PS with SCs significantly augmented the hyperthermia-induced YES1 activation during early recovery. In support of this observation, accumulated evidence has shown that SCs synthesize and secrete approximately 60 various proteins, among which many are known to act as general and/or tissue-specific survival factors (e.g., epidermal growth factor [EGF] and stem cell factor [SCF]) [41][42][43]. Furthermore, large volumes of data are indicative of an antiapoptotic effect of SC-GC interaction.…”

Section: Discussionmentioning

confidence: 79%

YES1 Activation Elicited by Heat Stress Is Anti-Apoptotic in Mouse Pachytene Spermatocytes1

Liang¹,

Dong²,

Zhao³

et al. 2013

Biology of Reproduction

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Deregulated expression of protein tyrosine phosphorylation has been implicated in testicular response to different stimuli. Herein, YES1, a nonreceptor protein tyrosine kinase, was found to be significantly up-regulated in pachytene spermatocytes (PS) during early recovery from a transient testicular heat stress. Coculture of PS with Sertoli cells (SCs) could enhance the hyperthermia-induced YES1 activation, indicative of a positive regulation of the paracrine signaling. Moreover, SU6656, a selective YES1 inhibitor, was shown to effectively block YES1 activity, thereafter resulting in a dramatic increase of heat stress-induced apoptosis in primary cultured PS. Mechanistically, the antiapoptotic effect of YES1 activation in response to testicular heat insult may mediate via the regulation of extracellular signal-regulated kinase (ERK)/metastasis-associated 1 (MTA1) cascade. From a clinical standpoint, a notably higher level of YES1 expression was observed in the pathological testis from varicocele patients as compared to a negligible staining in the control group. Taken together, our present results provide the first evidence that the YES1/ERK/MTA1/p53 cascade may serve as a naturally occurring, indispensable self-defensive mechanism maintaining apoptotic balance during meiotic heat stress. Our study may have also partially answered the question of how activation of signal pathways at the cell membrane surface interacts with the key regulatory events occurring in the nucleus during testicular heat shock.

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Section: Discussionmentioning

confidence: 79%

YES1 Activation Elicited by Heat Stress Is Anti-Apoptotic in Mouse Pachytene Spermatocytes1

Liang¹,

Dong²,

Zhao³

et al. 2013

Biology of Reproduction

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“…Without this factor, either spermatogonial aggregates do not develop or SSCs perish. 15 The generation process of SSCs and spermatogonia 9,16,17 and also the localization of undifferentiated spermatogonia along specific portions of the basement membrane are done via stimuli from the vascular network and interstitial cells, ie, the peritubular myoid cells and the Leydig cells. 18 On the other hand, the adhesion molecules of the basement membrane are anchored with SSCs.…”

Section: Introductionmentioning

confidence: 99%

The effects of poly L-lactic acid nanofiber scaffold on mouse spermatogonial stem cell culture

Eslahi¹,

Hadjighassem²,

Joghataei³

et al. 2013

IJN

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Introduction A 3D-nanofiber scaffold acts in a similar way to the extracellular matrix (ECM)/basement membrane that enhances the proliferation and self-renewal of stem cells. The goal of the present study was to investigate the effects of a poly L-lactic acid (PLLA) nanofiber scaffold on frozen-thawed neonate mouse spermatogonial stem cells (SSCs) and testis tissues. Methods The isolated spermatogonial cells were divided into six culture groups: (1) fresh spermatogonial cells, (2) fresh spermatogonial cells seeded onto PLLA, (3) frozen-thawed spermatogonial cells, (4) frozen-thawed spermatogonial cells seeded onto PLLA, (5) spermatogonial cells obtained from frozen-thawed testis tissue, and (6) spermatogonial cells obtained from frozen-thawed testis tissue seeded onto PLLA. Spermatogonial cells and testis fragments were cryopreserved and cultured for 3 weeks. Cluster assay was performed during the culture. The presence of spermatogonial cells in the culture was determined by a reverse transcriptase polymerase chain reaction for spermatogonial markers ( Oct4, GFRα-1, PLZF, Mvh(VASA), Itgα6 , and Itgβ1 ), as well as the ultrastructural study of cell clusters and SSCs transplantation to a recipient azoospermic mouse. The significance of the data was analyzed using the repeated measures and analysis of variance. Results The findings indicated that the spermatogonial cells seeded on PLLA significantly increased in vitro spermatogonial cell cluster formations in comparison with the control groups (culture of SSCs not seeded on PLLA) ( P ≤0.001). The viability rate for the frozen cells after thawing was 63.00% ± 3.56%. This number decreased significantly (40.00% ± 0.82%) in spermatogonial cells obtained from the frozen-thawed testis tissue. Both groups, however, showed in vitro cluster formation. Although the expression of spermatogonial markers was maintained after 3 weeks of culture, there was a significant downregulation for some spermatogonial genes in the experimental groups compared with those of the control groups. Furthermore, transplantation assay and transmission electron microscopy studies suggested the presence of SSCs among the cultured cells. Conclusion Although PLLA can increase the in vitro cluster formation of neonate fresh and frozen-thawed spermatogonial cells, it may also cause them to differentiate during cultivation. The study therefore has implications for SSCs proliferation and germ cell differentiation in vitro.

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“…Among the growth factors, GDNF (Glial-Cell-Line-Derived Neurotrophic Factor) is the most crucial factor to promote SSC self-renewal leading to a balance set between SSCs self-renewal and differentiation [47][48][49]. Without GDNF, spermatogonial aggregation cannot occur or SSCs may be perished [50]. In the cell culture, we experienced increased number of SSCs via self-renewal in presence of GDNF and somatic cells.…”

Section: Discussionmentioning

confidence: 96%

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

Azizollahi

Aflatoonian

Gilani

et al. 2016

J Assist Reprod Genet

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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.

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In vitroculture of testicular germ cells: Regulatory factors and limitations

Cited by 50 publications

References 157 publications

YES1 Activation Elicited by Heat Stress Is Anti-Apoptotic in Mouse Pachytene Spermatocytes1

YES1 Activation Elicited by Heat Stress Is Anti-Apoptotic in Mouse Pachytene Spermatocytes1

The effects of poly L-lactic acid nanofiber scaffold on mouse spermatogonial stem cell culture

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

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