Human oocyte maturation is considered as the reinitiation and completion of the first meiotic division from the germinal vesicle stage (prophase I) to metaphase II, and the accompanying cytoplasmic maturation for fertilization and early embryonic development. Immature human oocytes obtained from patients undergoing gynaecological surgery, or ovulation induction or having polycystic ovary syndrome (PCOS) can be matured and fertilized in vitro. To date, 80% of immature oocytes matured to metaphase II when cultured in maturation medium supplemented with gonadotrophins and 85% of matured oocytes fertilized and cleaved in vitro. Following transfer of these embryos, pregnancies and live births have been achieved. However, the capacity for oocyte maturation was different when the immature oocytes were retrieved from PCOS patients and when the oocytes were cryopreserved at germinal vesicle stage.
BackgroundTo investigate the effect of pectinase-treated Panax ginseng (GINST) in cellular and male subfertility animal models.MethodsHydrogen peroxide (H2O2)-induced mouse spermatocyte GC-2spd cells were used as an in vitro model. Cell viability was measured using MTT assay. For the in vivo study, GINST (200 mg/kg) mixed with a regular pellet diet was administered orally for 4 mo, and the changes in the mRNA and protein expression level of antioxidative and spermatogenic genes in young and aged control rats were compared using real-time reverse transcription polymerase chain reaction and western blotting.ResultsGINST treatment (50 μg/mL, 100 μg/mL, and 200 μg/mL) significantly (p < 0.05) inhibited the H2O2-induced (200 μM) cytotoxicity in GC-2spd cells. Furthermore, GINST (50 μg/mL and 100 μg/mL) significantly (p < 0.05) ameliorated the H2O2-induced decrease in the expression level of antioxidant enzymes (peroxiredoxin 3 and 4, glutathione S-transferase m5, and glutathione peroxidase 4), spermatogenesis-related protein such as inhibin-α, and specific sex hormone receptors (androgen receptor, luteinizing hormone receptor, and follicle-stimulating hormone receptor) in GC-2spd cells. Similarly, the altered expression level of the above mentioned genes and of spermatogenesis-related nectin-2 and cAMP response element-binding protein in aged rat testes was ameliorated with GINST (200 mg/kg) treatment. Taken together, GINST attenuated H2O2-induced oxidative stress in GC-2 cells and modulated the expression of antioxidant-related genes and of spermatogenic-related proteins and sex hormone receptors in aged rats.ConclusionGINST may be a potential natural agent for the protection against or treatment of oxidative stress-induced male subfertility and aging-induced male subfertility.
Age-related male sexual dysfunction covers a wide variety of issues, together with spermatogenic and testicular impairment. In the present work, the effects of cordycepin (COR), an active constituent of a nutrient powerhouse Cordyceps militaris Linn, on senile testicular dysfunction in rats was investigated. The sperm kinematics, antioxidant enzymes, spermatogenic factors, sex hormone receptors, histone deacetylating sirtuin 1 (SIRT1), and autophagy-related mammalian target of rapamycin complex 1 (mTORC1) expression in aged rat testes were evaluated. Sprague Dawley rats were divided into young control (2-month-old; YC), aged control (12-month-old; AC), and aged plus COR-treated groups (5 (COR-5), 10 (COR-10), and 20 (COR-20) mg/kg). The AC group showed reduced sperm kinematics and altered testicular histomorphology compared with the YC group (p < 0.05). However, compared with the AC group, the COR-treated group exhibited improved sperm motility, progressiveness, and average path/straight line velocity (p < 0.05–0.01). Alterations in spermatogenesis-related protein and mRNA expression were significantly ameliorated (p < 0.05) in the COR-20 group compared with the AC group. The altered histone deacetylating SIRT1 and autophagy-related mTORC1 molecular expression in aged rats were restored in the COR-20 group (p < 0.05). In conclusion, the results suggest that COR holds immense nutritional potential and therapeutic value in ameliorating age-related male sexual dysfunctions.
Immature human follicular oocytes were collected from unstimulated ovaries, matured and fertilized
in vitro and then transferred to patients with no ovarian dysfunction such as premature ovarian failure.
From 11 1 consenting donors, 422 immature oocytes were collected from 97 ovaries between January
1990 and October 1991. The number of oocytes collected from ovaries and their development were
recorded so that comparisons could be made among donors of different ages and ovarian condition,
such as menstrual cycle, cyclic and non-cyclic ovaries. The rate of fertilization in vitro showed a peak
in the 31-40-year age group; however, there was no statistical difference in the rate of oocyte maturation
and cleavage among the donors in the different age groups. Immature oocytes of the luted phase had
a significantly higher maturation rate than those of the follicular phase. There was no significant
difference in the number of recovered oocytes, or in the development of immature follicular oocytes,
between cyclic and non-cyclic ovaries. Mature follicular fluid and peritoneal fluid had a significant
effect on the development of immature follicular oocytes. Also, it was found that fertilized eggs
cleaved more frequently in the medium containing hypoxanthine compared with the medium without
hypoxanthine. Finally, from 21 transfer cycles, viable embryos were derived from immature follicular
oocytes, resulting in two pregnancies, both leading to the birth of normal babies. These findings
suggest that culture in vitro of immature follicular oocytes, from unstimulated ovaries, to a suitable
condition, could be used optimally for clinical applications such as human ovum donation
programmes.
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