ABSTRACT
Background: Spermatogonial stem cells are the foundation of spermatogenesis and male fertility. So, their maintenance and culture are very important
Objective: In this study, we assessed protective effects of the Calligonum on in vitro viability and apoptotic and antiapoptotic genes expression of spermatogonial stem cells
Materials and Methods: After 24 hr of culture, the spermatogonial stem cells were treated with 30 MuM dose of H[2]O[2] and then 10 Mug/ml the Calligonum extract was added for 3 wks. Viability was assessed by Trypan blue, apoptosis using PI-Annexin and finally Bax, Bcl-2 and P53 genes expression by Real-Time Polymerase chain reaction
Results: After 3 wk of treatment, viability in the Calligonum extract+H[2]O[2] group was significantly higher than H2O2 group alone [p=0.001]. In the Calligonum extract+H2O2 group, apoptosis, as well as expression of apoptotic genes [Bax and P53], was significantly lower than the group treated with H[2]O[2] alone
Conclusion: The results of this study showed that 30 MuM H[2]O[2] increased apoptosis but decreased viability in spermatogonial stem cells. Calligonum has antioxidant properties that can reduce apoptosis, Bax and P53 expression and increase the viability and Bcl-2 expression
ABSTRACT
Introduction: Nowadays, spermatogonial stem cells [SSCs] cultivation has been used by many researchers as an effective tool for infertility treatments. Oxidative conditions can be effective on cell proliferation and differentiation of these cells. So, the aim of this study was to establish oxidative stress model for antioxidant activity of some drugs investigation during SSCs in vitro culture
Methods: Neonatal NMRI male mice [3-5 day] were used for isolation of SSCs. The cell suspension was prepared by twice enzymatic digestion. The cell suspension contents were spermatogonial and sertoli cells and treated by different doses of H2O2 logarithmic concentrations from 0-100 and muM after 24 hours. To access the optimal dose, extra doses from 10-100 and muM was evaluated. After 2 hours of H2O2 treatment, viability was determined by Trypan blue assay. The data were analyzed using SPSS software and One-way ANOVA test
Results: Our data showed that spermatogonial stem cells colonies appeared after 4 days of isolation. These cells expressed OCT4 and PLZF proteins. Many of spermatogonial stem cells were removed after using higher doses of H2O2. The results showed that 30 and muM concentration of H2O2 could induce oxidative stress in spermatogonial stem cell during in vitro culture
Conclusion: According to this study, 30 and muM concentration of H2O2 can cause cell death lower than 50% of total number of cells and can increase oxidative stress in cultivation of SSCs. This model is a suitable tool for studying some new antioxidant drugs
ABSTRACT
There are often duplicated ureter with types of congenital anomalies and associated clinical complications. The unilateral duplicated ureter was observed as an incidence result in usual dissection. During routine dissection of the body of an adult male with middle-aged 40-50 year old, the duality of unilateral incompletely ureter was seen in above. Two branches of ureter are coalesce in 7 cm distally hilum of kidney and form a unit ureter. Right kidney and ureter were entirely normal. This variation was accruing because ureter bud split incompletely during embryogenesis. Kidney with bifid ureter forms a clear diagnosis of renal failure. These anatomical differences may exhibit pathological obstruction urinary tract and should be considered in endourological method. Awareness of relationship between renal arteries and collecting systems for applications is necessary and vital in endourological procedures and renal surgeries