Cryopreservation of spermatogonial stem cells (SSCs) is an applicable method for young males seeking fertility preservation before starting a treatment. It increases reactive oxygen species (ROS) formation and oxidative stress, which damages cellular structures. In this study, we added two antioxidants, catalase and α-tocopherol (α-TCP), to the basic freezing medium to evaluate their effects on the efficiency of SSCs. SSCs were isolated from testes of 3- to 6-day-old male mice using enzymatic digestion. The enrichment of isolated cells was evaluated by flow cytometry and Stra8 antibody. Catalase (40 μg/mL), or α-TCP (200 μg/mL) was added to the basic freezing medium. The cell viability was evaluated by the methylthiazoltetrazolium (MTT) assay. After thawing, cells were cultured for 1 month, and the expression pattern of specific genes of SSCs and the ability of the cells to restore spermatogenesis were used to determine the efficiency of the cryopreservation method. The survival rate of the frozen cells in the presence of catalase or α-TCP was significantly higher than the control group (p < 0.05). The number of colonies and their diameter measured after 1 month were significantly higher in the antioxidant groups than in the control group (p < 0.05). Gene expression and resumption of spermatogenesis also followed the same pattern. Thus, adding antioxidants to the basic freezing medium can be helpful in increasing the quality and viability of SSCs after cryopreservation. This new approach to stem cells cryopreservation can also be a promising strategy for fertility preservation in patients who suffer from malignancy.

Background:Cadmium (Cd) is known to cause various disorders in the testis. The general population may be exposed to Cd through ingestion of food and drinking water, inhalation of particulates from ambient air, tobacco smoke and ingestion of contaminated soil and dust. Saffron (Crocus sativus L.) is widely used as a food flavour, and has well known medicinal effects.Objectives:The aim of this study was to determine the effects of saffron on the results of semen parameters (sperm concentration, motility and viability in cauda of epididymis) in rats exposed to cadmium.Materials and Methods:Thirty Wistar male rats were divided into four groups. Cadmium chloride (1 mg/kg body weight) was injected intraperitoneally during 16 days at intervals of 48 hours between subsequent treatments. Crocus sativus L. (100 mg/kg b.w., IP) was pretreated in both control and cadmium-injected rats. Both control and cadmium-injected rats were pretreated with Crocus sativus L. (100 mg/kg b.w., IP). The animals were killed and their sperm count, motility, and vitality were evaluated.Results:Sperm parameters did not differ significantly between control and sham groups. Following contamination with cadmium, sperm count, motility and vitality were significantly decreased in comparison to control group (P < 0.05). In pretreated (saffron) group, the sperm parameters improved significantly in comparison with cadmium group (P ≤ 0.05). A significant decrease in sperm motility was observed in Cd-treated rats compared to the control rats. However, no significant changes were recorded by comparison of the control and saffron treated groups except for the sperm motility parameter.Conclusions:Saffron, as an antioxidant, is positively effective on sperm parameters in rats exposed with cadmium.

Activation and overexpression of FoxO3 in granulosa cells of PCOS women correlated with higher apoptosis levels in these cells suggesting that FoxO3 may be a candidate for the higher apoptosis in granulosa cells from women with PCOS.

Many studies have reported that human endometrial mesenchymal stem cells (HuMenSCs) are capable of repairing damaged tissues. The aim of the present study was to investigate the effects of HuMenSCs transplantation as a treatment modality in premature ovarian failure (POF) associated with chemotherapy‐induced ovarian damage. HuMenSCs were isolated from menstrual blood samples of five women. After the in vitro culture of HuMenSCs, purity of the cells was assessed by cytometry using CD44, CD90, CD34, and CD45 FITC conjugate antibody. Twenty‐four female Wistar rats were randomly divided into four groups: negative control, positive control, sham, and treatment groups. The rat models of POF used in our study were established by injecting busulfan intraperitoneally into the rats during the first estrus cycle. HuMenSCs were transplanted by injection via the tail vein into the POF‐induced rats. Four weeks after POF induction, ovaries were collected and the levels of Amh, Fst, and Fshr expression in the granulosa cell (GC) layer, as well as plasma estradiol (E2) and progesterone (P4) levels were evaluated. Moreover, migration and localization of DiI‐labeled HuMenSCs were detected, and the labeled cells were found to be localized in GCs layer of immature follicles. In addition to DiI‐labelled HuMenSCs tracking, increased levels of expression of Amh and Fshr and Fst, and the high plasma levels of E2 and P4 confirmed that HuMenSC transplantation had a significant effect on follicle formation and ovulation in the treatment group compared with the negative control (POF) group.

The findings of this study suggest that AG improves sperm DNA fragmentation that is associated with infertility in varicocelized rats, and treatment with AG can reduce the damage to sperm DNA.

Cryopreservation of spermatogonial stem cells is considered as a useful procedure for preserving fertility in children with testis cancer. SSCs were isolated from testes mice, and then antioxidant was added to the freezing medium. The Bax expression level in antioxidant groups was significantly (P ≤ 0.05) lower than the control group and a significant rise of Bcl2 expression was detected in the antioxidant groups. ROS production with antioxidant was significantly lower compared with the control group. Cryopreservation with the addition of the antioxidants can help increase the number of SSCs and improve the quality and viability of these cells after cryopreservation.

Manganese inhibits oxidative stress damage. The aim of this study was to investigate the protective role of manganese on testis structure and sperm parameters in adult mice exposed to formaldehyde (FA). Twenty adult male NMRI mice were selected and randomly divided into four groups: (i) control; (ii) sham; (iii) 'FA'-exposed group; and (iv) 'FA and manganese chloride'-exposed group. The FA-exposed groups received 10 mg kg(-1) FA daily for 14 days, and manganese chloride was just injected intraperitoneally 5 mg kg(-1) on 2nd weeks. Mice were sacrificed, and spermatozoa were collected from the cauda of the right epididymis and analysed for count, motility, morphology and viability. The other testicular tissues were weighed and prepared for histological examination upon removal. Seminiferous tubules, lumen diameters and epithelium thickness were also measured. The findings revealed that FA significantly reduced the testicular weight, sperm count, motility, viability and normal morphology compared with control group (P ≤ 0.05). In addition, seminiferous tubules atrophied and seminiferous epithelial cells disintegrated in the FA group in comparison with the control group (P ≤ 0.05). However, manganese improved the testicular structure and sperm parameters in FA-treated mice testes (P ≤ 0.05). According to the results, manganese may improve and protect mice epididymal sperm parameters and testis structure treated with FA respectively.

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