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Background: Effective culture systems for attachment, migration, proliferation, and differentiation of spermatogonial stem cells (SSCs) can be a promising therapeutic modality for preserving male fertility. Decellularized extracellular matrix (ECM) from native testis tissue creates a local microenvironment for testicular cell culture. Furthermore, platelet-rich plasma (PRP) contains various growth factors for the proliferation and differentiation of SSCs. Methods:In this study, human testicular cells were isolated and cultured for 4 weeks, and SSCs were characterized using immunocytochemistry (ICC) and flow cytometry. Human testicular tissue was decellularized (0.3% SDS, 1% Triton), and the efficiency of the decellularization process was confirmed by histological staining and DNA content analysis. SSCs were cultured on the human decellularized testicular matrix (DTM) for 4 weeks. The viability and the expression of differentiation genes were evaluated by MTT and real-time polymerase chain reaction (PCR), respectively.Results: Histological evaluation and DNA content analysis showed that the components of ECM were preserved during decellularization. Our results showed that after 4 weeks of culture, the expression levels of BAX, BCL-2, PLZF, and SCP3 were unchanged, while the expression of PRM2 significantly increased in the cells cultured on DTM supplemented with PRP (ECM-PRP). In addition, the expression of GFRA1 was significantly decreased in the ECM group compared to the control and PRP groups. Furthermore, the MTT test indicated that viability was significantly enhanced in cells plated on DTM supplemented with PRP. Conclusion:Our study demonstrated that DTM supplemented with PRP can provide an effective culture system for the differentiation and viability of SSCs.

Objectives: Trichomonas infection is prevalent in the United States and a metronidazole oral tablet is the medication of choice for treating this infection. Based on various side-effects of oral or vaginal metronidazole and the increase in microbial resistance against chemical antibiotics, the use of herbal medicine with fewer side-effects seems to be essential. Laboratory experiments indicate the strong anti-microbial effects of Prangos ferulacea (PF) medicinal herb. However, no clinical trial has focused on its anti-microbial effects in humans. Thus, the present study aimed to determine the effects of PF vaginal cream on accelerating Trichomonas vaginalis infection (TVI) recovery. Materials and Methods: The present randomized clinical trial was conducted on 80 non-pregnant women visiting the healthcare centers affiliated with Lorestan University of Medical Sciences, Iran, in 2018. Trichomonas infection was diagnosed based on patient complaints, clinical observations, as well as wet mount and stained microscopic tests. The women were randomly divided into two groups of 40 each. One group received oral metronidazole plus PF vaginal cream while the other received oral metronidazole plus placebo vaginal cream for 7 days. Clinical observations, along with wet mount and stained microscopic tests were performed during 7 days following the treatment. Finally, data were analyzed using independent-samples t test, as well as chi-square, Fisher exact, Mann-Whitney U, and McNemar tests at the significance level of P<0.05. Results: Based on the results, the response to treatment with oral metronidazole plus PF vaginal cream was 92.50 based on patient complaints. More precisely, 86.25% was based on clinical criteria (i.e., strawberry cervix, foamy greenish-yellow vaginal discharge, pH ≥4.5, and positive amine test) and 85% was related to the microscopic criteria of wet mount (x40) and polymorphonuclear leukocytes (x100). In addition, the response to treatment with oral metronidazole and placebo vaginal cream was 91.25%, 83.12%, and 80% based on the patient complaint, clinical criteria, and microscopic criteria, respectively. Eventually, the analysis of the patient complaint, clinical criteria, and microscopic criteria in each group revealed a significant difference before and after the treatment (P<0.001) Conclusions: The results of this study showed that the PF herbal vaginal cream can be used for the treatment of TVI as an effective treatment along with oral metronidazole.

Background Spermatogenesis refers to the differentiation of spermatogonial stem cells (SSCs) located in the basal part of seminiferous tubules into haploid spermatozoa. Prerequisites for in vitro spermatogenesis include an extracellular matrix, a culture medium containing factors influencing the survival, proliferation and differentiation of spermatogenic cells, and testicular somatic cells that play a supporting role for SSCs. Accordingly, in this paper, the potential of co-culture of Sertoli cells and SSCs embedded in a hybrid hydrogel of agarose and laminin as one of the main components of the extracellular matrix of the basal compartment of seminiferous tubules in the presence of culture medium containing growth factors was investigated. Methods After three–week conventional culture of the cellular suspension of enzymatic digestion of human testicular tissue, the cells were cultured in agarose hydrogel alone or hybrid hydrogel (agarose/laminin) for 74 days. Then, the presence of spermatogonial cells and other cells of the different stages of spermatogenesis were analysed by immunocytochemistry, real-time PCR, electron microscopy and morphological staining. Results The results showed that colonies with positive spermatogenesis markers were observed in both culture systems. The presence of different levels of spermatogenesis in two groups media was confirmed, but morphological spermatozoa were observed only in hybrid hydrogel group. Conclusions Based on the results of this paper, it can be concluded that an agarose and extracellular matrix-based scaffold which maintains the mechanical strength of the scaffold during culture and also increases the biological activity of the scaffold, can support from all physiological activities of SSCs such as survival, proliferation and differentiation.