Induce Oocyte Maturation Research Articles

Severe ovarian hyperstimulation syndrome with GnRH agonist trigger during the COVID pandemic: lessons learned from an unusual case

Background: Injectable gonadotropins stimulate multi-follicular recruitment and allows retrieval of multiple oocytes for assisted reproduction. The widespread utilization of gonadotropin releasing hormone agonist (GnRHa) to induce oocyte maturation for oocyte retrieval has nearly eliminated the risk of severe ovarian hyperstimulation syndrome (OHSS), and only a few cases have been reported in the literature. The rarity of severe OHSS may lead to the mistaken conclusion that gonadotropin stimulation can be safely administered with limited monitoring, even in high-risk patients. We present an unusual case of a woman with limited monitoring due to the COVID pandemic who developed severe OHSS before GnRH agonist trigger and oocyte. Case Presentation: A 29-year-old nulliparous woman with polycystic ovarian syndrome (PCOS) initiated ovarian stimulation for oocyte retrieval. She had a robust initial response, and developed worsening abdominal pain, bloating, nausea, vomiting, and decreased appetite before retrieval. GnRH agonist was given to “trigger ovulation and retrieval scheduled due to the low reported incidence of severe OHSS. Symptoms progressed, and on the morning of retrieval, ultrasound demonstrated bilaterally enlarged ovaries >10cm and 48 oocytes were retrieved for a planned cryo-all cycle. She was hospitalized on the day of retrieval for severe OHSS and had two large-volume paracenteses. She was stable and discharged home by day 5, and symptoms markedly improved with the onset of menses. She has an ongoing pregnancy from her first frozen embryo transfer. Conclusion: We add a rare case of severe OHSS with a GnRHa trigger and cryo-all protocol with the onset of symptoms before GnRH agonist administration. Although rare, severe OHSS may still occur with a GnRHa trigger, and caution is needed when an initial robust response is identified. Here we also provide an opportunity to review the important patient risk factors for the development of OHSS and measures to reduce the risk in excessive responders.

Vasopressin Promoter Transgenic and Vasopressin Gene-Edited Ascidian, Ciona intestinalis Type A (Ciona robusta): Innervation, Gene Expression Profiles, and Phenotypes.

Oxytocin (OT) and vasopressin (VP) superfamily neuropeptides are distributed in not only vertebrates but also diverse invertebrates. However, no VPergic innervation of invertebrates has ever been documented. In the ascidian, Ciona intestinalis Type A (Ciona robusta), an OT/VP superfamily peptide was identified, and the Ciona vasopressin (CiVP) induces oocyte maturation and ovulation. In the present study, we characterize the innervation and phenotypes of genetically modified Ciona: CiVP promoter-Venus transgenic and CiVP mutants. CiVP promoter-Venus transgenic Ciona demonstrated that CiVP gene was highly expressed in the cerebral ganglion and several nerves. Fluorescence was also detected in the ovary of young CiVP promoter-Venus transgenic ascidians, suggesting that the CiVP gene is also expressed temporarily in the ovary of young ascidians. Furthermore, a marked decrease of post-vitellogenic (stage III) follicles was observed in the ovary of CiVP mutants, whereas pre-vitellogenic (stage I) and vitellogenic (stage II) follicles were increased in the mutant ovary, compared with that of wildtype Ciona. Gene expression profiles showed that the expression of various genes, including genes related to ovarian follicle growth, was altered in the ovary of CiVP mutants. Altogether, these results indicated that CiVP, mainly as a neuropeptide, plays pivotal roles in diverse biological functions, including growth of early-stage ovarian follicles via regulation of the expression of a wide variety of genes. This is the first report describing a VP gene promoter-transgenic and VP gene-edited invertebrate and also on its gene expression profiles and phenotypes.

Investigation on the interrelation between total length, age,sex maturation and sex reversal in <italic>Epinephelus fasciatus</italic>

The growth, development and maturation are comprehensively affected by age, total length, sex hormone and so on in teleosts. The blacktip grouper <italic>Epinephelus fasciatus</italic> is an important and valuable marine economic species widely distributed in the coastline of east Asian countries. Recently, with the global warming and increase of sea water temperature in the coastline of Japan, the time of growth, development and maturation of this species has become shorter than before while the size of growth and maturation has become smaller than before. Therefore, the access to information and research work on the growth, development and maturation of this species becomes very important and indispensable in the coastline. To understand the interrelation between total length(TL), age, sex maturation, and sex reversal, <italic>Epinephelus fasciatus</italic> was captured in the coastline of Nagasaki city in Japan from July 2015 to June 2018. The TL, body weight, age, sex, gonadal developmental maturity, and gonadosomatic index (GSI) were measured and calculated. In addition, the changes of 17β-Estradiol (E₂) content in the serum were analyzed. The results indicated that the mature proportion and GSI of female <italic>Epinephelus fasciatus</italic> individuals reached the peak in July, suggesting that the sexual mature timing of this species might be in July. The mature female <italic>Epinephelus fasciatus</italic> individuals were firstly detected at 3 years old, and the TL was about 24.5 cm. Accompanied with the increase of TL, the quantities of the mature individuals gradually increased, which indicated that the age of the first sex maturation was at 3 years old in this species. In addition, at 3 and 4 years old in female <italic>Epinephelus fasciatus</italic>, the E₂ content in the mature individuals was significantly higher than that in the immature individuals (<italic>P</italic><0.05), suggesting that E₂ played an important role in inducing oocyte maturation in this species. The intersex individuals were detected at about 35.7 cm and 9 years old, and the male individuals were detected at about 36.7 cm and 7 to 12 years old. In summary, the sexual mature timing of the <italic>Epinephelus fasciatus</italic> in the coastline of Nagasaki city in Japan was in July, which was closely related to the increase of E₂ content. The first sex maturation and sex reversal were at 3 years old or 7 to 12 years old, respectively. Based on the investigation of <italic>Epinephelus fasciatus</italic> in the coastline of Nagasaki city, the interrelation between total length, age, sex maturation and sex reversal was studied to provide the basic biological data and theoretical basis for resource conservation and artificial propagation.

A relaxin-like gonad-stimulating peptide identified from the starfish Astropecten scoparius.

A relaxin-like gonad-stimulating peptide (RGP) in starfish was the first identified invertebrate gonadotropin responsible for final gamete maturation. An RGP ortholog was newly identified from Astropecten scoparius of the order Paxillosida. The A. scoparius RGP (AscRGP) precursor is encoded by a 354 base pair open reading frame and is a 118 amino acid (aa) protein consisting of a signal peptide (26 aa), B-chain (21 aa), C-peptide (47 aa), and A-chain (24 aa). There are three putative processing sites (Lys-Arg) between the B-chain and C-peptide, between the C-peptide and A-chain, and within the C-peptide. This structural organization revealed that the mature AscRGP is composed of A- and B-chains with two interchain disulfide bonds and one intrachain disulfide bond. The C-terminal residues of the B-chain are Gln-Gly-Arg, which is a potential substrate for formation of an amidated C-terminal Gln residue. Non-amidated (AscRGP-GR) and amidated (AscRGP-NH2 ) peptides were chemically synthesized and their effect on gamete shedding activity was examined using A. scoparius ovaries. Both AscRGP-GR and AscRGP-NH2 induced oocyte maturation and ovulation in similar dose-dependent manners. This is the first report on a C-terminally amidated functional RGP. Collectively, these results suggest that AscRGP-GR and AscRGP-NH2 act as a natural gonadotropic hormone in A. scoparius.

Transcriptome analysis of common carp (Cyprinus carpio) provides insights into the ovarian maturation related genes and pathways in response to LHRH-A and dopamine inhibitors induction

Luteinizing hormone-releasing hormone analog (LHRH-A) and dopamine inhibitors have been widely used to induce oocyte maturation and ovulation in domesticated fishes. Although this approach represents a reliable method for regulating fish reproduction, the underlying molecular mechanisms mediating LH action are largely unexplored. The objective of this study was to determine the transcriptional profile of gene programming in hormone-treated common carp. In the present study, female common carp were intraperitoneally injected with LHRH-A together with dopamine inhibitors, and control fish were injected with saline. Ovarian morphological changes were analysed by both light microscopy and scanning electron microscopy. Furthermore, gene expression profiling of the brain and ovarian tissues was performed by Illumina sequencing. Compared to the control carp, hormone treatment resulted in morphological changes including disappearance of nuclear membrane, breakdown of germinal vesicle (GVBD), and fusion of yolk globules, reflecting that hormones significantly promoted oocyte maturation. In comparison to control, we have identified 867 and 9,053 differentially expressed genes in the hormone-treated female brain and ovary, respectively. In the brain, most of the identified genes were significantly enriched in 18 KEGG pathways. In the ovarian tissue, the identified genes were significantly involved in 9 pathways. In the hormone-treated carp, genes were involved in calcium signalling pathway, cAMP signalling pathway, insulin secretion, and oxidative phosphorylation pathway, which showed obvious associations with ovarian maturation. The present study provides transcriptomic information for hormone-treated carp, which might be useful for studying the endocrine regulation and mechanisms of ovarian maturation in domesticated fishes.

Handling and Manipulation of Gametes and Embryos of the Annelidan Worm Pseudopotamilla occelata.

Pseudopotamilla occelata is a polychaete worm distributed widely in the northern part of the Pacific coast, having value as fishing bait as well as biological material for some basic research areas, including reproduction. Here we describe methods for handling the gametes and embryos of this worm, focusing on such topics as maintenance of adults, induction of oocyte maturation and fertilization, culture of embryos and larvae, microinjection into oocytes, and calcium (Ca2+) imaging.

Comparison of oocyte maturity rates in recombinant Human Chorionic Gonadotropin (HCG) and triptorelin acetate triggers: A prospective randomized study

Luteinizing Hormone (LH) like exposure in the mid cycle for inducing the oocyte maturation is the very crucial step in the success of ICSI treatment. Introduction of LH surge endogenously by GnRH-agonist for final oocyte maturation induction, may be more physiological compared with the administration of HCG. Since GnRH agonist would induce FSH surge also along with LH surge, as happens in natural cycle. However, the effects of giving HCG trigger for inducing only LH surge and giving GnRH agonist trigger for inducing both LH and FSH surge, in patients treated for ICSI with GnRH antagonists need more research. Sub fertile patients planned for ICSI, meeting the requirement of inclusion criteria, were started with recombinant FSH from day 2 of menstrual cycle. GnRH antagonists were started from day 6 of stimulation. FSH dose was adjusted according to the individual response. Trigger was planned when the lead follicle reaches 24 mm. For triggering, 100 patients were randomized to receive Recombinant HCG trigger and Triptorelin acetate trigger. Oocyte retrieval was done 36 hours after Recombinant hCG Trigger and 35 hours after Triptorelin acetate trigger. The oocyte maturity rate was assessed by the number of metaphase II oocytes retrieved.

Necessity is the mother of invention and the evolutionary force driving the success of in vitro fertilization.

During the last few decades, millions of healthy children have been born with the aid of in vitro fertilization (IVF). This success belies the fact that IVF treatment is comprised of a complex series of interventions starting with a customized control ovarian stimulation protocol. This is followed by the induction of oocyte maturation, the retrieval of mature oocytes and in vitro fertilization, which often involves the microinjection of a single sperm into the oocyte. After fertilization, the resulting embryos are cultured for up to 7 days. The best embryos are transferred into the uterus where the embryo implants and hopefully develops into a healthy child. However, frequently the best embryos are biopsied and frozen. The biopsied cells are analyzed to identify those embryos without chromosomal abnormalities. These embryos are eventually thawed and transferred with pregnancy rates as good if not better than embryos that are not biopsied and transferred in a fresh cycle. Thus, IVF treatment requires the coordinated efforts of physicians, nurses, molecular biologists and embryologists to conduct each of these multifaceted phases in a seamless and flawless manner. Even though complex, IVF treatment may seem routine today, but it was not always the case. In this review the evolution of human IVF is presented as a series of innovations that resolved a technical hurdle in one component of IVF while creating challenges that eventually lead to the next major advancement. This step-by-step evolution in the treatment of human infertility is recounted in this review.

Being on the side of old findings: progesterone elevation on the day of oocyte maturation induction does not affect embryological parameters throughout the blastocyst culture period.

To investigate whether there is any detrimental effect of progesterone elevation (PE) on the day of oocyte maturation induction on embryological development potentials. This retrospective single-center cohort study included a total of 1485 individual intracytoplasmic sperm injection (ICSI) cycles between January 2014 and December 2018. Serum progesterone (P) levels were measured on the day of oocyte maturation induction following the GnRH antagonist suppression protocol. Embryological parameters such as maturation, fertilization rate (FR), top-quality embryo (TQE) formation rate per 2PN on day 3, and excellent-quality blastocyst (EQB) formation rate per 2PN on day 5/6 were recorded. The inclusion criteria for women were an age ≤ 37years, a BMI ≤ 30kg/m2, and access to a total sperm concentration ≥ 2 million. Groups were stratified according to the serum P levels using the cut-off levels of < 0.8ng/ml; 0.8-1.49ng/ml; and ≥ 1.5ng/ml. Peak E2 level and total number of oocytes retrieved were significantly related to PE (p < 0.001). FR did not display a significance difference between groups (p = 0.108). The TQE and the blastulation rates were not affected by PE (p = 0.82 and p = 0.68, respectively). Chi square analysis revealed a significant relationship between PE and the EQB formation rate (p = 0.01). GEE analysis failed to present any statistical significance regarding the effect of PE on neither the TQE nor the EQB formation rates per 2PN [OR 1.07; 95% (0.98-1.16) p = 0.113 and OR 0.93; 95% (0.80-1.07) p = 0.32, respectively]. In accordance with previously published papers, our study could not find any detrimental effect of PE on embryological outcomes throughout the blastocyst culture period.

An agonist for membrane progestin receptor (mPR) induces oocyte maturation and ovulation in zebrafish in vivo

The maturation and ovulation of fish oocytes are well-characterized biological processes induced by progestins via coordination of nongenomic actions and genomic actions. Previously, we established a procedure that enables the induction of oocyte maturation and ovulation in live zebrafish by simple administration of the natural teleost maturation-inducing hormone 17 alpha, 20 beta-dihydroxy-4-pregnen-3-one (17,20β-DHP) into the surrounding water. By this in vivo assay, the potencies of chemicals in inducing or preventing oocyte maturation and ovulation can be evaluated. The potencies of compounds in inducing ovulation of zebrafish oocytes also can be evaluated in vivo with improved in vitro assays. Here, we attempted to evaluate the effect of Org OD 02–0 (Org OD 02), a selective agonist for membrane progestin receptor (mPR), on fish oocyte maturation and ovulation with in vitro and in vivo assays. As reported previously, Org OD 02 triggered oocyte maturation in vitro. The same Org OD 02 triggered oocyte maturation within several hours in vivo. Surprisingly, Org OD 02 even induced ovulation both in in vivo and in vitro. Eggs from Org OD 02-induced ovulation could be fertilized by artificial insemination. The juveniles developed normally. These results indicated that Org OD 02 triggered physiological ovulation in live zebrafish. In summary, we have demonstrated the effect of Org OD 02 on fish oocyte maturation and ovulation in vitro and in vivo. The results suggested that Org OD 02 acted as an agonist not only of mPR but also of nuclear progesterone receptor (nPR).

Metabolic changes in follicular fluids of patients treated with recombinant versus urinary human chorionic gonadotropin for triggering ovulation in assisted reproductive technologies: a metabolomics pilot study.

The main goal of this retrospective cohort study is the assessment of the effects of administration of recombinant-hCG (r-hCG) versus urinary-hCG (u-hCG) on follicular fluid (FF) composition of women who underwent in vitro fertilization (IVF) treatments. We selected 70 patients with infertility attributable to tubal diseases, unexplained infertility, and male factor. Metabolomics analysis of their FFs was performed by 1H nuclear magnetic resonance (1H NMR) spectroscopy in combination with multivariate analysis to interpret the spectral data. Univariate statistical analysis was applied to investigate the possible correlations between clinical parameters and between clinical parameters and metabolites identified by NMR. According to the type of hCG used, significant differences were detected in FFs of women with male factor and unexplained infertility, both in qualitative and quantitative terms, for some metabolites as cholesterol, citrate, creatine, β-hydroxybutyrate, glycerol, lipids, amino acids (Glu, Gln, His, Val, Lys) and glucose. No significant difference was observed in women with tubal diseases. Besides, the number of MII oocytes in the u-hCG-treated groups correlates positively with glutamate in tubal disease and with glycerol in unexplained infertility. In the r-hCG-treated groups, the number of MII oocytes correlates positively with lipid in tubal disease, positively with citrate and negatively with glucose in male infertility. Metabolite composition of FF changes according to different type of hCG treatment and this can be related to oocyte development and subsequent outcome. According to the data of this study, different types of hCG should be used in relation to the diagnosis of infertility to obtain better results in inducing oocyte maturation in women undergoing IVF.

Impaired oocyte maturation and ovulation in membrane progestin receptor (mPR) knockouts in zebrafish

Accumulating evidence suggest that membrane progestin receptor α (mPRα) is the membrane receptor mediating nongenomic progestin signaling that induces oocyte maturation in teleost. However, the involvement of other members of mPR family in oocyte maturation is still unclear. In this study, we found impaired oocyte maturation in zebrafish lacking mPRα1, mPRα2, mPRβ, or mPRγ2. In contrast, no difference was observed in oocyte maturation in the single knockout of mPRγ1, mPRδ, or mPRε. To study possible redundant functions of different mPRs in oocyte maturation, we generated a zebrafish line lacking all seven kinds of mPRs (mprs−/−). We found oocyte maturation was further impaired in mprs−/−. In addition, oocyte ovulation delay was observed in mprs−/− females, which was associated with low levels of nuclear progestin receptor (Pgr), a key regulator for ovulation. We also found reduced fertility in mprs−/− female zebrafish. Furthermore, eggs spawned by mprs−/− females were of poor quality.

Nodal regulates ovarian functions in zebrafish

Nodal, a member of the transforming growth factor-β (TGF-β) superfamily, plays critical roles during embryo development. Several studies suggest that Nodal also regulates reproduction. The objective of this study was to investigate if Nodal is expressed in zebrafish ovary and if it is involved in the regulation of ovarian functions. Using real-time PCR, we detected two Nodal homologs, nodal-related (ndr)1, and ndr2 in zebrafish ovarian follicles. We further compared the mRNA levels of ndr1, ndr2, and their receptors between maturational incompetent early vitellogenic follicles (stage IIIa) and mid- to late-vitellogenic follicles (stage IIIb) which are capable of undergoing maturation when they are induced by hormones. We found that mRNAs for ndr1 and ndr2, as well as a type I receptor, acvr1ba, were significantly increased in follicular cells isolated from stage IIIb follicles. In primary cultures of ovarian follicular cells, treatment with recombinant human Nodal inhibited cell proliferation. On the other hand, Nodal increased the mRNA levels of two steroidogenic enzymes hsd3b2 and cyp17a1, as well as paqr8, which encodes the membrane progestin receptor-β (mPR-β). Conversely, knockdown of ndr1 and ndr2 using siRNAs decreased the mRNA levels of hsd3b2, cyp17a1, and paqr8. Finally, treatment of Nodal significantly induced oocyte maturation. Taken together, these findings suggest that Nodal exerts multiple effects on zebrafish ovary to regulate follicle growth, steroidogenesis, and oocyte maturation.

Calling activity and calls' temporal features inform about fish reproductive condition and spawning in three cultured Sciaenidae species

Although many fish species are vocal, the use of fish sounds for aquaculture management and wild population protection has not received much attention. In this study, sound production of three members of the Sciaenidae family was monitored before and during spawning in aquaculture facilities. The species examined include the meagre Argyrosomus regius, the shi dum Umbrina cirrosa and red drum Sciaenops ocellatus. Red drum reproduces spontaneously in captivity, whereas shi drum and meagre often require hormonal stimulation using gonadotropin releasing hormone agonist (GnRHa) for the induction of oocyte maturation, ovulation and spawning. In all three species, a clear increase in calling activity was detected during spawning nights, when longer sounds with a higher number of faster repeated pulses were emitted. Changes of call temporal features could be related to histological changes in the sonic muscles. After GnRHa treatment and during spawning, meagre sonic fibres were wider and the ratio of sarcoplasmic reticulum to myofibril cross-sectional area was higher. The correlation of calling activity with spawning events opens up the possibility of using the monitoring of calling activity and of call temporal features as tools for evaluating the reproductive state of different sciaenid species, both in the wild and captivity.

Activin A overexpression promotes rat follicular development through SCF-kit-mediated cell signals

We explored the possible signaling pathway by which activin A induces oocyte maturation. Inhba-overexpressing lentivirus vectors were constructed and transfected into primary granulosa cells in vitro and ovary tissues in vivo in rats. The granulosa cell growth curve was drawn, and antibodies for phospho-Smad2, phospho-Erk5, phospho-Nur77, and stem cell factor (SCF) were prepared for western blot analysis. Protein expression of SCF and C-kit in the rat ovaries was detected by immunohistochemical staining. The rate of granulosa cell proliferation was higher in the Inhba gene overexpression group (INH) than in the control groups (CON group and GFP group) in vitro. Protein expression of SCF and C-kit was higher in the INH group than in the other two groups. phospho-Smad2, phospho-ERK5, P-nur77, and SCF proteins showed positive expression in rat ovarian granulosa cells in each group and were obviously increased in the INH group. Activin A overexpression may promote rat granulosa cell proliferation through Smad2/ERK5/nur77 signaling pathways, and rat granulosa cells overexpressing activin A in vitro showed increased levels of SCF and c-kit proteins.

Induction of oocyte maturation and changes in the biochemical composition, physiology and molecular biology of oocytes during maturation and hydration in the orange-spotted grouper (Epinephelus coioides)

In aquaculture, oocyte maturation, ovulation and spawning are important for successful breeding and seed production. Most marine teleost fish can be induced to spawn regularly using intramuscular injections of human chorionic gonadotropin (HCG). In this study, an intra-ovarian injection method was developed in orange-spotted grouper to deal with the broodstocks unresponsive to HCG. Successful oocyte maturation and ovulation in the unilateral injected ovary was induced within 35 h of treatment. An in vitro method was then established to induce HCG-primed oocyte maturation. HCG or maturation-inducing hormones (MIHs) induced the maturation of oocytes with a diameter between 490 and 560 μm. Biochemical, physiological and molecular changes during oocyte maturation and hydration were also characterized. The concentration of free amino acids except the cysteine increased during oocyte maturation, whereas Na+ decreased steadily and K+ increased. The expression of genes encoding cathepsin B, D, L, and the subunits of Na+, K+-ATPase were upregulated during hydration. Specific inhibitors for cathepsin and Na+, K+-ATPase were used to further confirm their crucial roles in oocyte hydration both in vivo and in vitro. These results provide valuable data for inducing oocyte maturation and evaluating oocyte quality in marine teleost fish.

Participation of Phosphatidylinositol-3 Kinase Signalling in Human Chorionic Gonadotropin, Bovine Insulin (B-Insulin) and Human-Insulin-Like Growth Factor-I Induced Oocyte Maturation and Steroidogenesis in the Grey Mullet, Mugil Cephalus.

The grey mullet, Mugil cephalus, is an edible fish of high economic importance. Breeding biology with reference to hormonal/growth factor regulation of oocyte maturation needs to be known for its commercial production. The present study was conducted to examine the potency of maturation inducing hormones, chorionic gonadotropin (hCG), bovine-insulin, and insulin like growth factor1 (h-IGF-1) I on ovarian steroidogenesis and oocyte maturation. The role of hormones and growth factors on steroidogenesis and oocyte maturation was investigated using specific inhibitors, Wortmannin for phosphatidylinositol-3 (PI3) kinase, trilostane for 3β-hydroxysteroid dehydrogenase, 1-octanol and 1-heptanol for gap junctions, actinomycin D for transcription and cycloheximide for translation of signal molecules. Actions of hormonal and growth factors were examined for steroidogenesis, by radioimmunoassay and oocyte maturation by germinal vesicle breakdown (GVBD). Specific inhibitors were used to determine the cell signaling pathways, PI3 kinase. All the inhibitors attenuated the hCG-induced oocyte maturation (GVBD%), steroidogenesis including transcription, translation, gap junctions and PI3 kinase signaling. These inhibitors failed to inhibit h-IGF-I and b-insulin-induced oocyte maturation, steroidogenesis, translation and PI3 kinase signaling. hCG induces oocyte maturation via steroid dependent pathway involving gap junctions, transcription, translation and PI3 kinase signaling, unlike h-IGF-I and b-insulin in the mullet.

In Vitro Generation of Oocytes from Ovarian Stem Cells (OSCs): In Search of Major Evidence.

The existence of ovarian stem cells (OSCs) in women as well as their physiological role in post-menopausal age are disputed. However, accumulating evidence demonstrated that, besides the animal models including primarily mice, even in adult women putative OSCs obtained from ovarian cortex are capable to differentiate in vitro into oocyte-like cells (OLCs) expressing molecular markers typical of terminal stage of oogonial cell lineage. Recent studies describe that, similarly to mature oocytes, the OSC-derived OLCs also contain haploid karyotype. As proof of concept of their stem commitment, OSCs from mice differentiated to oocytes in vitro are suitable to be fertilized and implanted in sterilized animals resulting in embryo development. Despite enthusiasm for these data, which definitely require extended confirmation before considering potential application in humans for treatment of ovarian insufficiency, OSCs appear suitable for other clinical uses, restoring the endocrine derangements in premature ovarian failure or for fertility preservation in oncologic patients after anti-cancer treatments. In this context, the selection of viable oocytes generated from OSCs before chemotherapy protocols would overcome the potential adjunct oncogenic risk in women bearing hormone-dependent tumors who are repeatedly stimulated with high dose estrogens to induce oocyte maturation for their egg recruitment and cryopreservation.

GnRH agonist versus HCG triggering in different IVF/ICSI cycles of same patients: a retrospective study

The aim of this study was to assess Gonadotropin Releasing Hormone agonist (GnRHa) trigger results of fresh in vitro fertilisation (IVF), Intracytoplasmic Sperm Injection (ICSI) cycles in high-responder patients. Thirty-six high-responder patients, undergoing GnRH antagonist protocol combined with GnRHa trigger for final oocyte maturation, were included. All cycles were autologous fresh transfer cycles. Fifteen of 36 patients had previous IVF/ICSI cycles triggered with human chorionic gonadotropin (hCG) and both cycles of these patients were compared. The mean fertilisation rate, blastocyst development and clinical pregnancy rates were 67%, 44.4% and 44.4%, respectively. The hCG and GnRHa trigger cycles of the same patients were compared as two groups (n: 15). 2PN oocyte counts were significantly higher in agonist trigger cycles (p .048). There were no differences in terms of M2 oocyte count and fertilisation rate. The blastocyst formation and clinical pregnancy rates for hCG and GnRHa trigger cycles were 33.3–66.7% and 13.3–46.7%, respectively. These results were found to be 2-fold and 3.5-fold higher, but not statistically significant. GnRHa trigger in combination with LPS is a good option for final oocyte maturation due to its good pregnancy outcomes and virtually eliminating OHSS risks.IMPACT STATEMENTWhat is already known on this subject? Gonadotrophin releasing hormone agonist (GnRHa) trigger is effective in the induction of oocyte maturation and prevention of Ovarian Hyperstimulation Syndrome (OHSS) on IVF cycles using antagonist protocol.What do the results of this study add? The main strength of this study is the comparison of different triggers in different cycles of the same patients. GnRHa trigger in combination with Luteal Phase Support (LPS) is a good option for final oocyte maturation due to its good pregnancy outcomes and virtually eliminating OHSS risks.What are the implications of these findings for clinical practice and/or further research? We suppose that GnRHa trigger combined with modified LPS is clinically more successful than Human Chorionic Gonadotropin (hCG) in regard to OHSS prevention and reproductive outcomes on fresh IVF/ICSI cycles. More extensive studies are needed to draw firm conclusions.

The potential application of conditioned media-mesenchymal stem cells on human oocyte maturation in assisted reproductive technology: a quasi-experimental based-study at Dr. Sardjito General Hospital, Yogyakarta, Indonesia

Background: PCOS typically characterized by a decreased level of GDF9 and BMP15, which contribute to follicular arrest. Recent research related to stem cells showed that conditioned media-mesenchymal stem cell (CM-MSC) might have a positive role in folliculogenesis. Therefore, this study aimed to assess the effectiveness of CM-MSC towards oocyte maturityMethod: A quasi-experimental design was used to examine the outcome of 3 different interventions, namely group A (4-hour incubation in standard media), group B (24-hour incubation in standard media), and group C (24 hours incubation in conventional media with addition of CM-MSC). Nuclear and cytoplasmic maturity, along with GDF9 and BMP15 levels were measured and analyzed.Results: Sixty-three patients at an infertility clinic, RSUP Dr. Sardjito, Yogyakarta was recruited and a total of 292 germinal vesicles were obtained to start in vitro maturation procedure. Multivariate analysis showed group C has OR 6.9 (3,3–14,41) to obtain metaphase II oocyte than group B (p &lt;0,0001). Infertility causes, insulin resistance, and maternal age risk are other factors that significantly influence oocyte maturity outcome (p &lt;0.001; p = 0.005; p = 0.017). For the oocyte morphology outcome, no significant effect was obtained from the intervention (p&gt; 0.05). Group C has higher GDF9 levels (Δ mean = 3.31) and BMP15 (Δ mean = 1.52) compared with group B (p &lt;0.001; p = 0.006).Conclusion: It can be concluded that 24-hour incubation in CM-MSC was effective to induce oocyte maturation. However, other factors, such as infertility causes, insulin resistance, and maternal age, should also be considered.