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Abstract
Simple SummaryMesenchymal stem cells and their derivatives are used in clinical studies for their anti-apoptotic, anti-oxidant, immunomodulatory, and regenerative properties. Their use in reproductive medicine is increasing as they have been proved to be beneficial for infertility treatment. Mesenchymal stem cells can secrete factors that influence biological processes in target tissues or cells; these factors are either directly secreted by the cells or mediated through their derivatives. Although the amniotic membrane is easy to obtain and is a good source of stem cells, clinical trials using amniotic membrane-derived mesenchymal stem cells are still uncommon, especially in reproductive medicine or artificial reproductive technologies. The objective of the present study was to demonstrate the effects of conditioned medium prepared from amniotic membrane-derived stem cells on dog sperm cryopreservation. Our results showed that 10% of the conditioned medium enhanced the quality-related parameters of frozen–thawed sperm cells because of the presence of antioxidants and growth factors in the medium, which probably protected spermatozoa during the freeze–thaw process. These results suggest that conditioned media prepared from amniotic membrane-derived mesenchymal stem cells might have clinical applications in assisted reproductive technologies.This study investigated the effects of conditioned medium (CM) from canine amniotic membrane-derived MSCs (cAMSCs) on dog sperm cryopreservation. For this purpose, flow cytometry analysis was performed to characterize cAMSCs. The CM prepared from cAMSCs was subjected to proteomic analysis for the identification of proteins present in the medium. Sperm samples were treated with freezing medium supplemented with 0%, 5%, 10%, and 15% of the CM, and kinetic parameters were evaluated after 4–6 h of chilling at 4 °C to select the best concentration before proceeding to cryopreservation. Quality-related parameters of frozen–thawed sperm were investigated, including motility; kinetic parameters; viability; integrity of the plasma membrane, chromatin, and acrosome; and mitochondrial activity. The results showed that 10% of the CM significantly enhanced motility, viability, mitochondrial activity, and membrane integrity (p < 0.05); however, the analysis of chromatin and acrosome integrity showed no significant differences between the treatment and control groups. Therefore, we concluded that the addition of 10% CM derived from cAMSC in the freezing medium protected dog sperm during the cryopreservation process.
Highlights
Sperm cryopreservation is used to store sperm samples from cancer patients or endangered species, and for other activities such as breeding, shipping, and research; the post-thaw quality obtained is low when compared with the fresh samples [1]
The percentage of spermatozoa with abnormal chromatin condensation-stained nuclei was not significantly different in both the groups (p < 0.05), (Table 4). These results suggest that canine amniotic membrane-derived Mesenchymal stem cells (MSCs) (cAMSCs)-conditioned medium (CM) has no protective effect on DNA integrity
Since the isolation of MSCs from amniotic tissues, researchers have displayed a growing interest in the study of their characteristics, properties, and possible applications [31,34,55]. They have become an interesting alternative to embryonic stem cells, as they can be obtained by non-invasive methods [31], and the ethical issues associated with the use of amniotic tissues-derived stem cells are minor [34], since they are obtained from the placenta that is usually discarded after caesarian sections [29]
Summary
Sperm cryopreservation is used to store sperm samples from cancer patients or endangered species, and for other activities such as breeding, shipping, and research; the post-thaw quality obtained is low when compared with the fresh samples [1]. Sperm cells are more sensitive to environmental changes because of their limited protein and lipid biosynthetic abilities [6] and the absence of DNA repair mechanism [7]. These events result in a weakened oxidative stress defense that exposes the sperm cells to reactive oxygen species (ROS). An increase in the ROS production during cryopreservation destroys sperm lipid matrix structures and, subsequently, causes the loss of membrane integrity, an increase in lipid peroxidation, and excessive DNA fragmentation [7,8]
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