ABSTRACT
Objective@#Optimizing culture media for the incubation of immature oocytes is a vital strategy to increase the oocyte maturation rate during in vitro maturation (IVM) programs. This study evaluated the IVM and fertilization rates of human germinal vesicle (GV) and metaphase I (MI) oocytes using two different maturation media (commercial and homemade) with or without growth differentiation factor 9-β (GDF9-β). supplementation. @*Methods@#Immature oocytes from intracytoplasmic sperm injection (ICSI) cycles were collected and assigned to one of two IVM culture media (commercial or homemade; cleavage-stage base). After maturation, MII oocytes were examined under an inverted microscope for the presence of the polar body, zona pellucida (ZP) birefringence, and meiotic spindle (MS) visualization after maturation in four conditions (commercial or homemade medium, with or without GDF9-β. ICSI was done for matured oocytes, and fertilization was confirmed by the visualization of two distinct pronuclei and two polar bodies. @*Results@#No significant differences were found between the two culture media in terms of the time and rate of oocyte maturation or the rate of fertilization (p>0.05). Growth factor supplementation increased the 24-hour maturation rate for both GV and MI oocytes only in homemade medium. The maturation rate after 24 hours was higher for MI oocytes (p0.05). @*Conclusion@#Higher rates of oocyte maturation and fertilization were observed after application of homemade medium supplemented with GDF9-β. Therefore, this combination may be recommended as an alternative for clinical IVM programs.
ABSTRACT
Background: Insulin resistance and hyperinsulinemia may play a role in pathogenesis of PCOS. One of the common therapeutic methods is using insulinsensitizing drugs such as metformin and thiazolidinediones
Objective: The purpose was to determine the effect of metformin and pioglitazone on clinical, hormonal and metabolic parameters in women with PCOS
Materials and Methods: Eighty four women randomly received one of the following for 3 months: metformin [n=28] [500 mg three times a day], pioglitazone [30 mg daily] [n=28] and combination of both metformin and pioglitazone [n=28] [30 mg/day pioglitazone plus 500 mg metformin three times a day]. Hormonal profile, fasting serum insulin, body weight, body mass index, menstrual status and waist to hip ratio were evaluated before and after treatment
Results: Metformin and pioglitazone and combination therapy induced favorable changes in fasting serum insulin, HOMA-IR index, QUICKI, fasting glucose to insulin ratio in women with PCOS. Body weight, BMI, and waist to hip ratio increased significantly after treatment with pioglitazone but the data were similar after administration of metformin or combination therapy. Total testosterone level decreased significantly only after treatment with metformin. After 3 months in patients who received pioglitazone or combination therapy, menstrual cycles became regular in 71.4% and 73.9% respectively. While menstrual improvement happened only in 36.4% of the patients treated with metformin
Conclusion: These findings suggest that insulin-sensitizing drugs induce beneficial effect in insulin resistance and menstrual cyclicity but only metformin ameliorated hyperandrogenemia in women with PCOS. Treatment with combination of metformin and pioglitazone did not show more benefit than monotherapy with each drug alone
ABSTRACT
Cyclophosphamide is a chemo-therapeutic agent used in the treatment of various cancers and autoimmune diseases. This composition has cytotoxic and clastogenic properties. The purpose of this study was to evaluate the protective effect of methanol extracts of Thymus vulgaris L. against DNA damage induced by cyclophosphamide in mouse bone marrow cells by the micronucleus test. The extract concentrations of 375, 750, 1500 mg/kg were injected intraperitoneally [Ip] into mice for 7 consecutive days. One hour after the last injection, cyclophosphamide 50 mg/kg Ip was injected. 24 hours after cyclophosphamide injection, the animals were killed and the samples of bone marrow were prepared and stained using the standard methods. For each sample, 1000 cells of polychromatic erythrocytes [PCE] and the same number of normochromatic erythrocyte [NCE] and the cells containing their micronucleus were counted. Cyclophosphamide increased the frequency of micronuclei polychromatic erythrocytes [MnPCE] and decreased cell proliferation [PCE/PCE+NCE]. All doses of extracts significantly reduced the micronucleus frequency ratio [P<0.05]. The cells proliferation ratio [PCE/PCE+NCE] was also increased. The best effect in reducing the micronucleus frequency was at 1500 mg/kg dosage. Thymus extract is able to reduce the clastogenic and cytotoxic effects of cyclophosphamide, due to its antioxidant properties, playing a protective role
ABSTRACT
In vitro maturation [IVM] of immature oocytes collected from ovary has been proposed for fertility preservation. In addition, quality of oocytes post IVM is one of the factors determining its developmental competence. By using the non-invasive Polscope system, both meiotic spindle [MS] and zona pellucida [ZP] can be assessed in living oocytes. The aim was to investigate the developmental potential of immature oocytes retrieved from ovarian tissue after IVM, as a method for fertility preservation, in patients with gynecological diseases. The ovarian cortex from 26 patients with malignant and benign diseases [21-45 years old], were obtained directly from collaborating hospitals, and transported to the IVF center on ice. In total 61 immature oocytes were aspirated, of which 18 [29.5%] were degenerated and discarded. The remaining 43 [70.5%] healthy oocytes were cultured in IVM culture media for 48 hr. The rate of maturity was assessed, and the ZP birefringence and MS were imaged with Polscope technology. Overall 43 immature oocytes underwent IVM technology, of which 30.2% reached viable metaphase II [MII] oocytes. The ovarian tissues of 9 [34.6%] women were lacking oocytes at any stage. During polarized light microscopy examination, MS could be visualized only in one of the MII oocytes, but high ZP birefringence's were observed in the majority of the oocytes post IVM [61.5%]. Oocytes maturation post IVM from unstimulated ovaries showed a good developmental competence in gynecologic patients. Further studies should be performed to advance the oocyte maturation program, such as co-culture system, for fertility preservation
ABSTRACT
In general, 15% of oocytes collected in ART cycles are immature. These oocytes may be cryopreserved further for use in in-vitro maturation [IVM] program. The aim of this study was to determine maturation capacity, morphometric parameters and morphology of human immature oocytes in both fresh IVM [fIVM] and vitrified-IVM [vIVM] oocytes. 93 women who underwent controlled ovarian stimulation for ART were included. The immature oocytes [n=203] were divided into two groups: the first group [n=101] directly matured in vitro; and the second group [n=102] first vitrified, then matured in vitro. All oocytes underwent IVM in Ham's F10 supplemented with LH+FSH and human follicular fluid. After 48h of incubation, the oocyte maturation rates, as well as morphometric and morphologic characteristics were assessed using cornus imaging and were compared. Oocyte maturation rates were reduced in vIVM, [40.4%], in comparison with fIVM [59.4%, p<0.001]. Following morphometric assessment, there was no difference in the mean oocyte diameters [micro m] between fIVM and vIVM, 156.3 +/- 6.8 and 154.07 +/- 9.9, respectively. Other parameters of perimeters, egg areas, as well as oocyte and ooplasm volumes were similar in two groups. In addition, more morphologic abnormalities, such as, vacuole, and dark oocyte were observed in vIVM oocytes. fIVM was more successful than vIVM groups. No statistical differences were noticed in morphometry assessment in two groups. This suggests that morphometric parameters can not be applied as prognosis factor in oocyte maturation outcome in IVM program