Protective effects of human amniotic membrane derived mesenchymal stem cells (hAMSCs) secreted factors on mouse spermatogenesis and sperm chromatin condensation following unilateral testicular torsion

Khadivi, Farnaz; Mojaverrostami, Sina; Ramesh, Mahya; Rastegar, Tayebeh; Abbasi, Yasaman; Bashiri, Zahra

doi:10.1016/j.aanat.2023.152084

Cited by 3 publications

(1 citation statement)

References 49 publications

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“…These results are not very different from those obtained in several studies on laboratory mouse strains. For example, we found a range of 0-0.5% darkly stained sperm with aniline blue in the three closely related mouse species, whereas laboratory mouse sperm exhibited ~5% of darkly stained sperm [59] although in some strains it reached 15-22% [57,58,60]. With methylene blue (as incorporated in the Diff-Quik kit), the three mouse species we examined showed 0.5-1.1% darkly stained spermatozoa; however, to the best of our knowledge, there are no studies using this dye to assess chromatin status in mouse sperm cells.…”

Section: Discussionmentioning

confidence: 81%

Sperm Chromatin Status and DNA Fragmentation in Mouse Species with Divergent Mating Systems

Agudo-Rios,

Sanchez-Rodriguez,

Idrovo

et al. 2023

IJMS

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Sperm DNA integrity and chromatin status serve as pivotal indicators of sperm quality, given their intricate link to sperm function, embryo development, and overall fertility. Defects in chromatin compaction, which are often associated with compromised protamine content, can lead to damaged DNA strands. In this study, the chromatin status and possible correlation with DNA damage was assessed in males of three mouse species: Mus musculus, M. spretus, and M. spicilegus. We employed various staining methods, including aniline blue, methylene blue (Diff-Quik), toluidine blue, and chromomycin A3, to assess chromatin compaction in cauda epididymal sperm. Samples were also analyzed by the sperm chromatin structure assay (SCSA) to estimate DNA fragmentation (%tDFI, %HDS). Analyses were carried out on freshly collected sperm and cells incubated for 3 h in a HEPES-buffered modified Tyrode’s medium simulating conditions of the female reproductive tract. Notably, the analysis of chromatin status yielded minimal abnormal values across all three species employing diverse methodologies. SCSA analyses revealed distinct variations in %tDFI between species. Following sperm incubation, the percentages of sperm stained with methylene blue exhibited differences among the species and were significantly correlated to the DNA fragmentation index. HDS demonstrated correlations with the percentages of sperm stained by aniline blue, methylene blue, and chromomycin A3. Overall, chromatin compaction was high across all species, with limited differences among them. The relationship between chromatin status and DNA integrity appeared to be related to levels of sperm competition among species.

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Section: Discussionmentioning

confidence: 81%

Sperm Chromatin Status and DNA Fragmentation in Mouse Species with Divergent Mating Systems

Agudo-Rios,

Sanchez-Rodriguez,

Idrovo

et al. 2023

IJMS

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Secretome of human amniotic membrane stem cells promote recovery and testicular functions through modulating SIRT1/NRF2/TNF-α pathway in mice testicular torsion: An experimental study

Esfehani,

Khadivi,

Valipour

et al. 2024

IJRM

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Background: Testicular ischemia/reperfusion injury, a significant result of testicular torsion, can lead to the risk of male infertility. Objective: The current study aimed to evaluate the effect of human amniotic membrane-derived mesenchymal stem cells (hAMSCs) secretome on testicular torsion/detorsion (T/D) in mice. Materials and Methods: All the experiments were performed in the Anatomy Department of Tehran University of Medical Sciences, Tehran, Iran, during the period of March 2023 to December 2023. 40 male NMRI mice (5–7 wk, 25–30 gr) were randomized into: 1) the sham group: mice received sham operations with no other interventions, 2) T/D group, 3) negative control group; torsion detorsion + intratesticular injection of Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12, and 4) the T/D group + hAMSCs secreted factors. Serum testosterone levels, hematoxylin and eosin staining, and sperm quality parameters were used to evaluate the therapeutic effects of hAMSCs secreted factors on the testicular structure and function. Tissue oxidative stress was measured by determining malondialdehyde, superoxide dismutase, catalase, and glutathione peroxidase-1. Nuclear factor erythroid 2-related factor 2, Kelch-like ECH-associated protein 1, NAD-dependent deacetylase sirtuin-1, tumor necrosis factor-alpha and tumor protein P53 mRNA expressions were assessed in testis via real-time polymerase chain reaction. Results: The results showed that hAMSCs secreted factors alleviated testicular T/D injury by attenuating oxidative stress, inflammatory response, and apoptosis via modulating the sirtuin-1/ nuclear factor erythroid 2-related factor 2 / tumor necrosis factor-alpha signaling pathway. Conclusion: hAMSCs secreted factors increased antioxidative, anti-inflammatory, and antiapoptotic properties which consequently increased testosterone levels, spermatogenesis, and sperm quality parameters.

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Sperm Chromatin Status and DNA Fragmentation in Mouse Species with Diverging Mating Systems

Agudo-Rios,

Sanchez-Rodriguez,

Idrovo

et al. 2023

Preprint

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Sperm DNA integrity and chromatin status serve as pivotal indicators of sperm quality, given their in-tricate link to sperm function, embryo development, and overall fertility. Defects in chromatin com-paction, which are often associated with compromised protamine content, can lead to damaged DNA strands. In this study, chromatin status and possible correlation with DNA damage was assessed in males of three mouse species: Mus musculus, M. spretus and M. spicilegus. We employed various staining methods, including aniline blue, methylene blue (Diff-Quik), toluidine blue and chromomy-cin A3, to assess chromatin compaction in cauda epididymal sperm. Samples were also analyzed by the sperm chromatin structure assay (SCSA) to estimate DNA fragmentation (%tDFI, %HDS). Analyses were carried out on freshly collected sperm and cells incubated for 3 h in a Hepes-buffered modified Tyrode's medium simulating conditions of the female reproductive tract. Notably, the analysis of chromatin status yielded minimal abnormal values across all three species, employing diverse meth-odologies. SCSA analyses revealed distinct variations in %tDFI between species. Following sperm incubation, the percentages of sperm stained with methylene blue exhibited differences among the species and were significantly correlated to the DNA fragmentation index. HDS demonstrated correla-tions with the percentages of sperm stained by aniline blue, methylene blue and chromomycin A3. Overall, chromatin compaction was high across all species, with limited differences among them. The relationship between chromatin status and DNA integrity appeared to be related to levels of sperm competition among species.

show abstract

Protective effects of human amniotic membrane derived mesenchymal stem cells (hAMSCs) secreted factors on mouse spermatogenesis and sperm chromatin condensation following unilateral testicular torsion

Cited by 3 publications

References 49 publications

Sperm Chromatin Status and DNA Fragmentation in Mouse Species with Divergent Mating Systems

Sperm Chromatin Status and DNA Fragmentation in Mouse Species with Divergent Mating Systems

Secretome of human amniotic membrane stem cells promote recovery and testicular functions through modulating SIRT1/NRF2/TNF-α pathway in mice testicular torsion: An experimental study

Sperm Chromatin Status and DNA Fragmentation in Mouse Species with Diverging Mating Systems

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