Effect of conditioned medium of umbilical cord-derived mesenchymal stem cells as a culture medium for human granulosa cells: An experimental study

Background: The umbilical cord-derived mesenchymal stem cells conditioned medium (UC-MSCs-CM) produces secretomes with anti-apoptotic properties, and has the potential to prevent apoptosis of granulosa cells (GC) during controlled ovarian hyperstimulation. Objective: To observe the effect of UC-MSCs-CM on the interaction between pro- and anti-apoptotic proteins and the influence of growth differentiation factor 9 (GDF9) production in GC. Materials and Methods: UC-MSCs-CM was collected from umbilical cord stem cell culture on passage 4. GC from 23 women who underwent in vitro fertilization were cultured and exposed to UC-MSCs-CM for 24 hr. Then RNA of the GC was extracted and the mRNA expression of BCL-2 associated X (BAX), survivin and GDF9 were analysed using quantitative real-time PCR. The spent culture media of the GC were collected for measurement of insulin growth factor 1 using ELISA. Results: The expression of BAX was significantly different after UC-MSCs-CM exposure (4.09E-7 vs. 3.74E-7, p = 0.02). No significant changes occurred in survivin, BAX/survivin ratio, and GDF9 expression after UC-MSCs-CM exposure (p > 0.05). The IGF-1 level of the CM was significantly higher after the CM was used as a culture medium for GC (2.28 vs. 3.07 ± 1.72, p ≤ 0.001). A significant positive correlation was found between survivin and GDF9 (r = 0.966, p ≤ 0.001). Conclusion: IGF-1 produced by UC-MSCs-CM can work in paracrine fashion through the IGF receptor, which can inhibit BAX and maintain GDF9 production. Moreover, under the influence of UC-MSCs-CM, GC are also capable of producing IGF-1, which can impact GC through autocrine processes. Key words: Conditioned medium, BAX, Survivin, GDF9, IGF-1.

A study on outcome of controlled ovarian hyperstimulation and intrauterine insemination (husband) in a medical college in Kolkata

Background: A combination of controlled ovarian hyperstimulation and intrauterine insemination (IUI) remains an important treatment option for couple having infertility. Success rate of IUI with ovulation induction ranges from 8-20% depending on many factors. Aims and Objectives: To assess the factors affecting the success rate of IUI and to evaluate the success of ovulation Induction by different methods of controlled stimulation protocol. Materials and Methods: It is a duration-based prospective cross-sectional study where total of 67 couples were included by inclusion and exclusion criteria. They underwent 90 cycles of IUI with each couple having a maximum of three cycles. Ovulation induction was done by clomiphene citrate or letrozole or gonadotrophins. Semen preparation was done by density gradient method. The outcomewas measured by positive urine pregnancy test. Range, percentage, confidence interval, mean with standard deviation, median, range, and P-value were calculated. P<0.05 was taken as statistically significant. Results: Out of 90 IUI cycles 8 were successful resulting in a success rate of 8.8% per cycle and 11.9% per couple. Factors that had a positive impact were follicle size >21 sqmm, endometrial thickness >9 mm, post wash count >15 million/ml, >2 cycles of IUI and on the number of follicles 2 or more on the day of trigger. Conclusion: The study concluded that IUI after ovulation induction can be a simple and safe cost-effective procedure in selected group of infertile couple. Clinical significance IUI following ovulation induction can be a successful approach for specific indications in a low-resource setting where options for other ART interventions are absent or limited.

Serum Sex Hormone Binding Globulin Concentration as a Predictor of Ovarian Response During Controlled Ovarian Hyperstimulation

Purpose: Serum concentrations of sex hormone binding globulin (SHBG), a glycated homodimeric plasma transport protein, correlate positively with the total number of follicles in women with infertility. However, the relationship between serum SHBG concentrations and the ovarian response during controlled ovarian hyperstimulation (COH) and whether this relationship differs between women with and without polycystic ovary syndrome (PCOS) remains unclear.Methods: The study cohort included 120 participants (60 non-PCOS and 60 PCOS) undergoing in vitro fertilization. Serum samples were collected from each participant every 2–3 days during the COH cycle. The concentrations of serum SHBG and other sex hormones were determined to investigate the relationship between serum SHBG concentrations and the ovarian response in women with and without PCOS.Results: We found that the serum SHBG concentration was positively correlated with the ovarian response in non-PCOS patients but not in PCOS patients.Conclusion: The serum SHBG concentration may be clinically useful as a predictor of the ovarian response during COH in patients without PCOS.

Controlled Ovarian Hyperstimulation Protocol in Infertile Patients During the COVID-19 Pandemic

Objectives: To explore the appropriate controlled ovarian hyperstimulation (COH) protocols in infertility patients who received the in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) treatments during the COVID-19 pandemic.Materials and Methods: This retrospective cohort study evaluated the efficiency of the early follicular-phase long-acting GnRH-agonist long (EFLL) protocol (a new protocol developed by Chinese clinicians), prolonged pituitary down-regulation of EFLL protocol (Pro-EFLL), and the GnRH-ant protocol for couples meeting the study criteria between February 2020 and June 2020 who were treated by the First Affiliated Hospital of Zhengzhou University during the COVID-19 pandemic, and compared the pregnancy rates and miscarriage rates per fresh transfer cycle, number of retrieved oocytes, endometrial thickness on the day of hCG injection and the number of fertilized oocytes, mature oocytes, fertilized oocytes, and transferable embryos among the three protocols.Results: We found that the prolonged pituitary down-regulation during the COVID-19 pandemic by utilizing a full-dose of GnRH-a administrated in infertility patients were no differences in clinical outcomes than other protocols, The prolonged pituitary down-regulation protocol and EFLL protocol were associated with a higher Endometrial thickness on the day of hCG injection (12.67 ± 2.21 vs. 12.09 ± 2.35 vs. 10.79 ± 2.38, P &lt; 0.001), retrieved oocytes (14.49 ± 6.30 vs. 15.02 ± 7.93 vs. 10.06 ± 7.63, P &lt; 0.001), mature oocytes (11.60 ± 5.71 vs. 11.96 ± 6.00 vs. 7.63 ± 6.50, P &lt; 0.001), fertilized oocytes (9.14 ± 5.43 vs. 8.44 ± 5.34 vs. 5.42 ± 5.20, P &lt; 0.001), and transferable embryos (4.87 ± 2.96 vs. 6.47 ± 5.12 vs. 3.00 ± 3.28 vs. P &lt; 0.001) in the GnRH-antagonist protocol.Conclusion: We recommend that patients start Gn injections 33–42 days after a pituitary downregulated full dose (3.75 mg) of gonadotropin-releasing hormone agonist during the COVID-19 pandemic, even a delay of 2–4 weeks does not affect the implantation rate. The study can provide a more detailed estimate and clinical management strategies for infertile couples during the COVID-19 pandemic.

Different Occurrence Rates of Centrally Located Cytoplasmic Granulation in One Cohort Oocytes Show Distinctive Embryo Competence and Clinical Outcomes

Background Centrally located cytoplasmic granulation (CLCG) is a common cytoplasmic dysmorphism in human oocytes retrieved after controlled ovarian hyperstimulation (COH). This study sought to achieve a better understanding of its formation and effects on clinical outcomes. Methods We retrospectively analyzed the data from 422 intracytoplasmic sperm injection (ICSI) treatment cycles. Three groups of patients were classified according to the rates of CLCG occurrence in one egg cohort, as partial (pCLCG) all (aCLCG) and no CLCG (control). Results The pCLCG group had a significantly lower Body Mass Index (BMI) and higher Anti-Mullerian hormone (AMH) level compared to the control or aCLCG groups. Consistent with these distinctive features in the pCLCG group, fertilization and blastocyst formation rates were reduced significantly in the pCLCG group but not in the aCLCG group. Furthermore, the clinical outcomes in fresh embryo transfer cycles were dramatically reduced in the pCLCG group compared with the control group. However, in frozen/thawed cycles, all three clinical outcomes were significantly reduced in the aCLCG group but not in the pCLCG group. Conclusion We propose that pCLCG may reflect a specific population of patients, and that the CLCG structure is sensitive to cryopreservation.

Maternal Antimüllerian Hormone as a Predictor of Euploid Is Independently Associated With Cumulative Live Birth Rates

Background: Serum antimÜllerian hormone (AMH) level has been reported to be associated with pregnancy rates after assisted reproduction. It is unclear whether AMH levels affect number of euploid blastocysts and cumulative live birth rates (CLBR). The aim of this study was to investigate whether AMH levels are associated with cumulative live birth rates (CLBR) through their relationship with oocyte quality.Methods: A total of 975 consecutive infertile women undergoing 1825 preimplantation genetic screening (PGS) analysis. Serum AMH levels were measured by AMH Gen II assay kit within the 3 months before Controlled Ovarian Hyperstimulation. Embryos were cultured and biopsied at the blastocyst stage. Results: Among 975 women undergoing PGS analysis. Age, serum AMH and number of oocytes retrieved were significantly and independently related to number of euploid blastocysts available for patients to transfer (P < .001). in patients displaying superior serum AMH levels, cumulative live birth rates (CLBR) were significantly increased independently from age and number of oocytes retrieved (P < .001), and this group with lower AMH levels also showed increased pregnancy loss rates.Conclusions: Serum AMH levels are positively age-independent associated with cumulative live birth rates after all viable embryos are transferred from the first ovarian stimulation cycle. And embryonic euploidy outcome was superior in patients with higher AMH levels. These present findings confirm that serum AMH levels might reflect not only ovarian reserve but also qualitative aspects of oocytes, which will effect the clinical application of PGS.