Evaluation of the expression level of microRNA-21, microRNA-15a, microRNA-372 in human follicular fluid stem cells-derived oocyte-like cells (OLCs).

OBJECTIVE: Today, researchers have succeeded in achieving oocyte-like cells through the in vitro differentiation of stem cells. MicroRNAs are key regulators of oocyte development. In this study we decided to evaluate the expression pattern of microRNA-21, microRNA-15a, and microRNA-372 in oocyte-like cells, to determine the maturation stage of oocyte-like cells. METHODS: Human follicular fluid samples were collected and centrifuged, and their cells were divided into 3 groups; day 7 as control group, days 14 and 21. During this period, the cells were evaluated for their morphological appearance and viability by inverted microscopy. RNA isolation was performed and cDNA was reversely transcribed by specific stem-loop RT primers. Real-time RT-PCR was used to detect microRNA expression. RESULTS: The relative expression of microRNA-21 and microRNA-15a on day 21 was significantly down-regulated compared to the control group (day 7), but microRNA-372 did not show a significant difference. Also, on day 14 compared to the control group (day 7), microRNA-21 did not show a significant difference; but microRNA-15a and microRNA-372 were significantly down-regulated. MicroRNA-21 and microRNA-15a on day 21 compared to day 14 revealed down-regulated levels, but microRNA-372 revealed up-regulated levels. CONCLUSIONS: Our results showed significant decreases in the expression of microRNA-21 and microRNA-15a in oocyte-like cells, as well as in oocytes, which may lead to cytoplasmic maturation, germinal vesicle break down and the completion of meiosis І. In addition, down-regulation expression of microRNA-372 maybe a confirmation that mesenchymal stem cells have differentiated into germ cells, and these cells were differentiated into oocyte-like cells.

Assisted Reproductive Techniques and submucous myoma.

Submucous myomas have negative effects on fertility. To maintain fertility, conservative treatment should be suggested to women who wish to become pregnant, especially young patients. The patient was a 33-year-old woman, who had had secondary infertility for 3 years. Upon vaginal ultrasound, we noticed a submucous myoma measuring 26 mm x 31 mm with a compressive effect on the anterior surface of the endometrium. Ovarian reserve was low. The gold standard of myoma treatment is surgical intervention. But, for the following reasons: the adverse effects of surgery on the endometrium (intrauterine adhesion), the patient's refusal to undergo a myomectomy and her request for pregnancy, our strategy for treating was to reduce volume of submucous myoma and start the assisted reproductive techniques (ART) cycle, simultaneously. We administered three courses of Gonadotropin-releasing hormone analogues (GnRHa) and then induced controlled ovarian hyperstimulation. Ovum pick up was done. Finally, we transferred two embryos (4 and 6 cells). In subsequent patient visits, ßhCG was positive after 14 days. At the last patient visit, the heart of the embryo was formed. From this finding, it may be concluded that combined GnRHa and ART is the treatment of choice for infertile women with uterine submucous myoma, considering the reduced ovarian reserve and response.

Effects of Vitamin D on Apoptosis and Quality of Sperm in Asthenozoospermia.

OBJECTIVE: Vitamin D receptor (VDR) is expressed in human spermatozoa. However, the role of vitamin D (VD) in human male reproduction has not yet been clarified. In this study, effects of VD on sperm parameters and its apoptosis in asthenozoospermic and healthy men were evaluated. METHODS: The study was carried out on discharged semen samples of 80 asthenozoospermic and healthy men. The samples were divided into control and experimental groups (received 20 µMol of VD). This study assessed sperm motility using the Makler chamber, their morphology by Diff quick, apoptosis and necrosis by Annexin-V and TUNEL assays, and their chromatin integrity was assessed by Aniline blue and Toluidine blue staining, according to WHO guidelines. RESULTS: The results revealed that: 1) the total number of motile sperms was increased by VD in both groups, but it was only significant in the asthenozoospermia group. 2) The progressive motility was increased with significant difference in both groups.3) Morphology of sperm did not show any changes due to VD in any of the groups. 4) Early apoptosis and necrosis of sperms were reduced in both groups, but the results of late apoptosis showed no statistical difference in these groups. 5) The percentage of positive toluidine blue was significantly decreased after using VD in the asthenozoospermia group. CONCLUSION: VD could improve motility, early apoptosis, and sperm necrosis, especially in asthenozoospermic men and it could be used for therapeutic opportunities.

A successful pregnancy following recurrent implantation failure with clinical laboratory strategy.

In treatment cycles of in vitro fertilization (IVF), 15% of the oocytes are immature and in the germinal vesicle (GV) phase. In rare occasions, more than 50% of the oocytes of a patient in a cycle are immature. Selecting fertility treatment for patients in this situation can be very challenging. The patient described in this report was a 35-year-old woman with primary infertility for 10 years. She had undergone fertility treatment six times, with no success. In her cycles, more than 50% of the oocytes were immature and in the GV phase. To address the situation, we developed a coordinated protocol involving medical and embryology procedures, analyzed the patient's medical records, and looked into the reasons of prior treatment failure. The development of a special and coordinated effort - instead of having medical and embryology teams working in isolation - proved efficient at providing better outcomes to the patient.