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A) Immature germinal vesicle (GV) oocytes isolated from 4-6 week-old mice 48 hr after injection with 5 IU pregnant mare serum gonadotropin (PMSG), enclosed with or without compact cumulus cells (Scale bar: 50 μm). B) GV oocytes at 24 hr of culture with distinct first polar body (Scale bar: 50 μm). C) Developed 2-cell and D) Blastocyst embryos obtained from fertilized GV oocytes cultured in potassium simplex optimization medium (KSOM) (Scale bar: 50 μm).
Source publication
Background: This study aimed to investigate the maturation and fertilization rates of immature mouse oocytes using Embryonic Stem Cell Conditioned Medium (ESCM).
Methods: Germinal Vesicle (GV) stage oocytes were observed in 120 NMRI mice, aged 4-6 weeks. GV oocytes with or without cumulus cells were subjected to IVM in either ESCM, Embryonic Stem C...
Contexts in source publication
Context 1
... GV oocytes were released from the ovaries by puncturing the follicles with a sterile 28-gauge needle as visualized under a stereomicroscope. The preantral follicles were pooled and randomly divided for further analysis ( Figure 1A). ...
Context 2
... those oocytes that dis- played distinct first polar bodies were classified as MII oocytes. The MII oocytes underwent fertilization and in vitro development ( Figure 1A). ...
Context 3
... medium is a conventional and chemically defined me- dium for development of mouse embryos. After insem- ination, the obtained 2pn embryos were cultured in KSOM under mineral oil at 37C in an atmosphere of 5% CO 2 and air for four days until the blastocyst stage ( Figure 1D ...
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The beneficial effects of resveratrol on in vitro maturation (IVM) have been explained mainly by indirect antioxidant effects and limited information is available on the underlying mechanism by which resveratrol acts directly on porcine cumulus oocyte complexes (COCs). Recently, several studies reported that sonic hedgehog (SHH) signaling mediates...
Citations
... Even by other researchers, in mice, the supplement of ESC conditioned medium in culture medium for in vitro maturation of oocytes were increased the development to blastocyst stage after fertilization, and these blastocysts were gave birth to normal healthy offspring after transplantation into surrogate mothers (Miraki et al., 2017). In pig, Kim and Park (2019) (Table 3 and 4). ...
... In assisted reproductive technology (ART), the maturation of immature oocytes in vitro is an essential technique for preserving female fertility [2]. In recent years, the development and activation of immature follicles have been the subject of numerous efforts, with the ultimate goal of treating infertility, premature ovarian failure, and polycystic ovary syndrome after cancer therapies [3,4]. ...
Oocyte cytoplasmic maturation is a crucial process during in vitro maturation (IVM), and finding an appropriate IVM medium that promotes oocyte competence is very critical in assisted reproductive technology (ART). The aim of this study was to investigate the effects of umbilical cord Wharton’s jelly (WJ-MSCs)-derived conditioned media on the maturation of immature oocytes and their developmental potential in humans after IVM, as well as apoptotic gene expression. A total of 392 germinal vesicle (GV) oocytes were collected from 207 women aged 25–35 years and divided into two IVM groups: (1) control group, which was cultured in CleavTM medium, and (2) experimental group, which was cultured in supernatants of umbilical cord Wharton’s jelly as a conditioned medium (CM). First, WJ-MSCs were isolated, and their purity was analyzed. The immunophenotypes of WJ-MSCs were analyzed by flow cytometry. The quantitative expression of BCL2, BAX, and BAG1 in matured oocytes and embryos was evaluated through quantitative real-time polymerase chain reaction (qRT-PCR). Our findings showed that WJ-MSCs have a high proliferating capacity. The purity of the isolated cells was further validated by immunophenotyping, which revealed that their surface antigen expression had phenotypical properties similar to WJ-MSCs. When compared to CD34 and CD45 surface markers, the enlarged cells were positive for CD90, CD105, and CD44. There were significant differences in cytoplasmic maturation of oocytes and embryo quality between the two groups. The mRNA expression levels of BCL-2, BAG1, and BAX in matured oocytes and embryos were also significantly different between the two groups. Therefore, WJ-MSCs medium indicated potential efficacy in terms of ameliorating oocyte maturation and in promoting the development and genes expression of BAX, BCL-2, and BAG1.
... Embryonic stem cells (ESCs) are pluripotent cells resulting from the inner cell mass (ICM) of preimplantation embryos in blastocyst stage (9,10). The ESCs can secrete biological products and activated proteins that HESCM for IVM subsequently provide an medium culture with mitogenic factors, growth factors, cytokines, and chemokines (11,12) which might be useful to improve IVM outcomes. ...
... Since many factors are involved in the maturation and development of oocytes, much research has been done based on the selection of more appropriate culture medium containing these factors (11)(12)(13). For example, factors such as EGFs have been shown to increase the rate of oocyte maturation (13). ...
... There are some factors secreted by ESCs (EGF, LIF, TGF-β, IGF) in the culture medium which supports IVM of oocytes (12). The effect of these factors on IVM, have been verified in several species (17,18). ...
Objective:
Some reports have indicated that conditioned medium from growing mouse embryonic stem cells (ESCs) provides a supportive condition for small follicles growing, oocyte maturation, and following embryo growth. The aim of this study is assessing in vitro maturation (IVM) and consequent in vitro fertilization (IVF) outcome of immature mouse oocytes using human embryonic stem cells conditioned medium (HESCM).
Materials and methods:
In this experimental study, 240 germinal vesicle (GV) oocytes were took from NMRI female mice, aged 4-6 weeks, 48 hours before injection of 5 IU pregnant mare serum gonadotropin (PMSG). 120 GV oocytes without cumulus cells were cultured in each of the groups. 120 GV were cultured in HESCM as test groups and also 120 GV cultured in human embryonic stem cells medium (HESM) as control groups. After evaluating the metaphase II (MII) oocyte maturation rate at 8, 16 and 24 hours, the MII oocytes subsequently were fertilized in vitro and the two-cell embryo development rate was recorded at days 1, 2, and 3. Statistical analysis was performed by using the generalized estimating equations (GEE) method that calculated their rate ratio.
Results:
Our data indicated there are significant differences between the maturation rates in HESCM and HESM (P=0.004), also the two-cell embryo development was significant between two culture media (P=0.00).
Conclusion:
Similar to some other studies, the secretome of the HESCM showed a significant impact on the IVM outcomes in mice.
... Moreover, it has been reported that the conditioned media A B Conditioned Media Improve IVM and IVF Efficiency obtained from embryonic stem cells (32), mesenchymal stem cells (16), and cumulus-oocyte-complexes (33) improve the rate of IVM in different species, suggesting that factors contained in such conditioned media regulate mechanisms involved in oocyte maturation. ...
Objective:
In vitro maturation (IVM) and cryopreservation of oocytes are two important parts of assisted reproductive technology (ART), but their efficacy is low. This study aimed to improve the quality of in vitro vitrified-warmed maturated oocytes using granulosa cell conditioned medium (GCCM).
Materials and methods:
In the experimental study, fresh/non-vitrified and vitrified-warmed mouse germinal vesicle (GV) oocytes (as F and V) were In vitro maturated using basal medium (BM) and also BM supplemented with 50% GCCM as treated groups (GM), and categorized as FBM, FGM, VBM and VGM groups, respectively. The rate of successful IVM (MII oocyte formation), mitochondrial membrane potential and the viability of MII oocytes were determined using inverted microscopy, JC-1 and trypan blue staining. Then, the rate of In vitro fertilization (IVF) and subsequent two-cell embryo formation was calculated. Finally, the expression levels of Oct4, Sox2, Cdk-2, Gdf9, Integrin beta1 and Igf2 were analyzed using real-time polymerase chain reaction (PCR) in MII oocytes and two-cell embryos.
Results:
These analyses showed that GCCM significantly increased the IVM rate, oocyte meiotic resumption and mitochondrial membrane potential (P<0.05). In addition, the rate of IVF and two-cell embryo formation was significantly higher in FGM and VGM compared to FBM and VBM (P<0.05). Interestingly, GCCM significantly affected the expression of the studied genes.
Conclusion:
Our findings suggest that GCCM might be useful for improving the efficiency of IVM and the subsequent IVF outcomes.
... The main challenge in IVM is the preparation of an adequate medium, which provides the most similar microenvironment to the in vivo condition (4,5). Recent studies have shown that the conditioned medium (CM) from mesenchymal and embryonic stem cells (MSCs and ESCs, respectively) used for IVM, can significantly improve the oocyte maturation and embryo development rates (6,7,8). ...
Background:
Testicular cell conditioned medium (TCCM) has been shown to induce female germ cell development In Vitro from embryonic stem cells (ESCs). Testicular cells (TCs) secrete a variety of growth factors such as growth differentiation factor-9 (GDF-9), bone morphogenetic protein 4 (BMP-4), stem cell factor (SCF), leukemia inhibitory factor (LIF), and other, that could improve oocyte maturation. Here we have investigated the effect of human TCCM (hTCCM) on in vitro maturation (IVM) and morphology of mouse oocytes.
Materials and methods:
In this experimental study, 360 germinal vesicle (GV) oocytes were obtained from NMRI mice, aged 4-6 weeks that had received 5 IU pregnant mare's serum gonadotropin (PMSG) 48 hours before. GV oocytes were subjected to IVM. 120 GV oocytes were cultured in each medium; hTCCM as the test group, DMEM + 20%FBS as the control group and Ham's F10 + HFF medium as the sham group. The rates of the IVM and perivitelline space (PVS) changes were recorded at 8, 16 and 24 hours after culture. The metaphase II (MII) oocytes were subjected for in vitro fertilization (IVF) and the fertilization rate was evaluated after 1, 2, and 3 days.
Results:
There was a significant difference between the maturation rates in hTCCM (31.67% MII) and the control [0% MII, P<0.05, (7.5% MI, 52.5% deg. and 40%GV)] groups but there was not a significant difference between the maturation rates in hTCCM and the sham group (53.33% MII, P>0.05). IVF success rate for MII oocytes obtained from IVM in the hTCCM group was 28.94% (n=11). Our data showed that hTCCM is an effective medium for GV oocyte growth and maturation compared to the control medium.
Conclusion:
Our findings show that TCCM supports oocyte IVM in mice and affect oocyte morphology.
The production of high-quality embryos in the laboratory and a successful pregnancy are closely related to the condition and contents of oocyte and embryo culture media. In this study, we investigated the effects of embryonic stem cell-conditioned medium (ESCCM) and embryonic stem cells growth medium (ESCGM) compared with potassium-enriched simplex optimized medium (KSOM) on preimplantation embryo development stages during natural or in vitro fertilization (IVF). Birth rate of pups was measured. To obtain mature oocytes, and 2-cell and 8-cell embryos, human chorionic gonadotropin (HCG) was injected 48 h after i.p. injection of 5 units of pregnant mare serum gonadotropin. Mature oocytes were obtained from non-mated female mice 14 h after HCG injection. To obtain 2-cell and 8-cell embryos, mated female mice, 1 day and 3 days, respectively, after HCG injection, were used. Mature oocytes were fertilized in HTF medium. Embryos obtained from natural or in vitro fertilization were cultured in experimental media, ESCCM and ESCGM, or KSOM as the control culture medium. Embryos that developed to the blastocyst stage were transferred to the uteri of pseudopregnant mice and effects of the experimental media on embryo viability were determined. ESCCM and ESCGM could not pass the embryo after the 2-cell stage, but they were suitable for the development of the embryo from the 8-cell stage to the blastocyst. It can be concluded that the embryo has various requirements at different stages of development.
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 <0,0001). Infertility causes, insulin resistance, and maternal age risk are other factors that significantly influence oocyte maturity outcome (p <0.001; p = 0.005; p = 0.017). For the oocyte morphology outcome, no significant effect was obtained from the intervention (p> 0.05). Group C has higher GDF9 levels (Δ mean = 3.31) and BMP15 (Δ mean = 1.52) compared with group B (p <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.
