scholarly journals Live offspring from vitrified blastocysts derived from fresh and cryopreserved ovarian tissue grafts of adult mice

Reproduction ◽  
2009 ◽  
Vol 138 (3) ◽  
pp. 527-535 ◽  
Author(s):  
Xiaoqian Wang ◽  
Sally Catt ◽  
Mulyoto Pangestu ◽  
Peter Temple-Smith
Keyword(s):  
Cancer Patients ◽  
Oocyte Maturation ◽  
Ovarian Tissue ◽  
Subsequent Development ◽  
Cumulus Cells ◽  
Ovarian Tissue Cryopreservation ◽  
Blastocyst Formation ◽  
Adult Mice ◽  
Tissue Grafts

Ovarian tissue cryopreservation and transplantation can be used to preserve fertility for cancer patients. In this study, we assessed the viability and function of ovarian tissue from adult mice that was cryopreserved by solid surface vitrification or traditional slow-cooling using variousin vitroandin vivotechniques, including allotransplantation,in vitrooocyte maturation, embryo culturein vitro, blastocyst cryopreservation, embryo transfer, and development. The importance of cumulus cells for oocyte maturation, fertilization, and embryo development was investigated. Graft recovery, follicle survival, and oocyte retrieval was similar in control, vitrified, and slow-cooled groups. High rates of oocyte maturation, cleavage, and blastocyst formation were achieved, with no significant differences between the control, vitrified or slow-cooled ovarian tissue grafts. The presence of cumulus cells was important for oocyte maturation, fertilization, and subsequent development. Cumulus–oocyte complexes with no surrounding cumulus cells (N-COCs) or with an incomplete layer (P-COCs) had significantly lower rates of oocyte maturation and blastocyst formation than cumulus–oocyte complexes with at least one complete layer of cumulus cells (F-COCs; maturation rate: 63, 78 vs 94%; blastocyst rate: 29, 49 vs 80%). Live births were achieved using vitrified blastocysts derived from oocytes taken from vitrified and slow-cooled ovarian tissue heterotypic allografts. Successful production of healthy offspring from these vitrified blastocysts suggests that this technique should be considered as a useful stage to pause in the assisted reproduction pathway. This provides an alternative protocol for restoring fertility and offering cancer patients a better indication of their chances of pregnancy and live birth.

scholarly journals Successful in vitro culture of pre-antral follicles derived from vitrified murine ovarian tissue: oocyte maturation, fertilization, and live births

Reproduction ◽  
2011 ◽  
Vol 141 (2) ◽  
pp. 183-191 ◽  
Author(s):  
Xiaoqian Wang ◽  
Sally Catt ◽  
Mulyoto Pangestu ◽  
Peter Temple-Smith
Keyword(s):  
Cancer Patients ◽  
Oocyte Maturation ◽  
Ovarian Tissue ◽  
Blastocyst Stage ◽  
Ovarian Tissue Cryopreservation ◽  
Antral Follicles ◽  
Live Births ◽  
And Function ◽  
Tissue Cryopreservation

Cryopreservation of ovarian tissue is an important option for preserving the fertility of cancer patients undergoing chemotherapy and radiotherapy. In this study, we examined the viability and function of oocytes derivedin vitrofrom pre-antral follicles as an alternative method for restoring fertility. Pre-antral follicles (specified as secondary follicle with a diameter around 100–130 μm) were mechanically isolated from vitrified-warmed and fresh adult mouse ovarian tissues and cultured for 12 days followed by an ovulation induction protocol at the end of this period to initiate oocyte maturation. Oocytes were then released from these follicles, fertilizedin vitro, and cultured to the blastocyst stage and vitrified. After storage in liquid nitrogen for 2 weeks, groups of vitrified blastocysts were warmed and transferred into pseudo-pregnant recipient females. Although most of the isolated mouse pre-antral follicles from fresh (79.4%) and vitrified (75.0%) ovarian tissues survived the 12-dayin vitroculture period, significantly fewer mature oocytes developed from vitrified-warmed pre-antral follicles than from the fresh controls (62.2 vs 86.4%,P<0.05). No difference was observed in embryo cleavage rates between these two groups, but the proportion of embryos that developed into blastocysts in the vitrification group was only half that of the controls (24.2 vs 47.2%,P<0.05). Nevertheless, live births of healthy normal pups were achieved after transfer of vitrified blastocysts derived from both experimental groups. This study shows that successful production of healthy offspring using anin vitrofollicle culture system is feasible, and suggests that this procedure could be used in cancer patients who wish to preserve their fertility using ovarian tissue cryopreservation.

scholarly journals Advances in the Treatment and Prevention of Chemotherapy-Induced Ovarian Toxicity

2020 ◽  
Vol 21 (20) ◽  
pp. 7792
Author(s):  
Hyun-Woong Cho ◽  
Sanghoon Lee ◽  
Kyung-Jin Min ◽  
Jin Hwa Hong ◽  
Jae Yun Song ◽  
...  
Keyword(s):  
Cancer Patients ◽  
Fertility Preservation ◽  
Ovarian Tissue ◽  
Alkylating Agents ◽  
Survival Rates ◽  
Chemotherapeutic Agents ◽  
Reproductive Age ◽  
Experimental Techniques ◽  
Ovarian Tissue Cryopreservation

Due to improvements in chemotherapeutic agents, cancer treatment efficacy and cancer patient survival rates have greatly improved, but unfortunately gonadal damage remains a major complication. Gonadotoxic chemotherapy, including alkylating agents during reproductive age, can lead to iatrogenic premature ovarian insufficiency (POI), and loss of fertility. In recent years, the demand for fertility preservation has increased dramatically among female cancer patients. Currently, embryo and oocyte cryopreservation are the only established options for fertility preservation in women. However, there is growing evidence for other experimental techniques including ovarian tissue cryopreservation, oocyte in vitro maturation, artificial ovaries, stem cell technologies, and ovarian suppression. To prevent fertility loss in women with cancer, individualized fertility preservation options including established and experimental techniques that take into consideration the patient’s age, marital status, chemotherapy regimen, and the possibility of treatment delay should be provided. In addition, effective multidisciplinary oncofertility strategies that involve a highly skilled and experienced oncofertility team consisting of medical oncologists, gynecologists, reproductive biologists, surgical oncologists, patient care coordinators, and research scientists are necessary to provide cancer patients with high-quality care.

scholarly journals Graft site and gonadotrophin stimulation influences the number and quality of oocytes from murine ovarian tissue grafts

Reproduction ◽  
2006 ◽  
Vol 131 (5) ◽  
pp. 851-859 ◽  
Author(s):  
Hsiao Yun Yang ◽  
Shae-Lee Cox ◽  
Graham Jenkin ◽  
Jock Findlay ◽  
Alan Trounson ◽  
...  
Keyword(s):  
Ovarian Tissue ◽  
Germinal Vesicle ◽  
Fertilization Rate ◽  
Ovarian Tissue Cryopreservation ◽  
Control Group ◽  
Developmental Potential ◽  
Oocyte Yield ◽  
Tissue Grafts

Ovarian tissue cryopreservation and subsequent transplantation can restore fertility in cancer patients. This study used a mouse ovarian grafting model to investigate whether the graft site (bursal cavity, the kidney capsule or subcutaneous) influences the number, fertilization rate and developmental potential of oocytes recovered from grafts and whether using a standard gonadotrophin stimulation protocol would increase oocyte yield from the grafts. Mouse ovarian tissue was grafted into four week old mice and collected three weeks later. Graft recipients were treated either with or without exogenous gonadotrophin stimulation prior to graft collection. Grafted ovaries yielded oocytes that were either at the germinal vesicle (GV) stage or mature metaphase II (MII) stage at collection. These GV oocytes were matured beforein vitrofertilization (IVF), while the MII oocytes underwent IVF immediately. Oocytes collected from the oviducts of non-grafted superovulated mice of the same age served as controls. Two-cell embryos were transferred to pseudopregnant recipients and recovered at day 15 of gestation or left to go to term. Graft retrieval and the number of oocytes from each graft were lowest from the subcutaneous graft site. The number of two-cell embryos produced was significantly higher for oocytes from the grafts to the bursa as compared with the other sites. All graft sites gave rise to embryos with comparable implantation rates and developmental potential to fetuses and offspring following transfer. However, the oocytes from grafted ovaries had a significantly lower developmental potential when compared with the control group. Stimulation with exogenous gonadotrophins did not significantly increase oocyte yield from grafted ovaries but did enhance oocyte maturation and development. In conclusion, graft site affects the number and quality of oocytes produced from ovarian grafts.

scholarly journals A Method for Chronological Intravital Imaging of Bovine Oocytes during In Vitro Maturation

2008 ◽  
Vol 14 (6) ◽  
pp. 549-560 ◽  
Author(s):  
Morten R. Petersen ◽  
Michael Hansen ◽  
Birthe Avery ◽  
Ingrid B. Bøgh
Keyword(s):  
Embryonic Development ◽  
Oocyte Maturation ◽  
Laser Scanning ◽  
Subsequent Development ◽  
Cumulus Cells ◽  
Fertilization Rate ◽  
Laser Scanning Microscopy ◽  
Intravital Imaging ◽  
Bovine Oocytes

AbstractOocyte maturation is known to affect the chances for successful fertilization, embryonic development, establishment of pregnancy and delivery of a live, healthy, and viable offspring. Two-photon laser scanning microscopy (TPLSM) has previously been used to evaluate early embryonic development without a detectable impairment of subsequent development, but has never been applied to assess mammalian oocytes throughout in vitro maturation (IVM). Visualization of structures within live oocytes during IVM, followed by fertilization and embryo culture, may improve the understanding of oocyte maturation. To visualize structures within bovine oocytes using TPLSM, it is necessary to remove the cumulus cells that normally surround the oocyte during maturation. Repeated visualization of structures within the same oocyte is possible, if movement of the oocyte can be avoided. In this article, we describe the development of a method for repeated intravital imaging of denuded bovine oocytes using an upright TPLSM equipped with a specially constructed incubator. Oocytes were stained with Hoechst 33258, and the nuclear structures were evaluated. Oocyte fertilization rate was not affected by TPLSM exposure, but the developmental capacity of the denuded oocytes was significantly reduced. This is, to our knowledge, the first article describing repeated intravital imaging during mammalian oocyte maturation using TPLSM.

228 EFFECT OF INSULIN ON BOVINE OOCYTE MATURATION, CUMULUS CELL APOPTOSIS, AND EARLY EMBRYO DEVELOPMENT IN VITRO

2015 ◽  
Vol 27 (1) ◽  
pp. 203
Author(s):  
I. Lindgren ◽  
P. Humblot ◽  
D. Laskowski ◽  
Y. Sjunnesson
Keyword(s):  
Embryo Development ◽  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Insulin Treatment ◽  
Cumulus Cells ◽  
Early Embryo ◽  
Blastocyst Formation ◽  
Early Embryo Development ◽  
Maturation In Vitro

Dairy cow fertility has decreased during the last decades, and much evidence indicates that metabolic disorders are an important part of this decline. Insulin is a key factor in the metabolic challenge during the transition period that coincides with the oocyte maturation and may therefore have an impact on the early embryo development. The aim of this study was to test the effect of insulin during oocyte maturation on early embryo development by adding insulin during the oocyte maturation in vitro. In this study, abattoir-derived bovine ovaries were used and cumulus-oocyte complexes (n = 991) were in vitro matured for 22 h according to standard protocols. Insulin was added during maturation in vitro as follows: H (10 µg mL–1 of insulin), L (0.1 µg mL–1 of insulin), or Z (0 µg mL–1 of insulin). After maturation, oocytes were removed and fixed in paraformaldehyde before staining. Click-it TUNEL assay (Invitrogen, Stockholm, Sweden) was used for apoptotic staining and DRAQ5 (BioNordika, Stockholm, Sweden) for nuclear staining (n = 132). Cumulus-oocyte complexes were evaluated using laser scanning confocal microscope (Zeiss LSM 510, Zeiss, Oberkochen, Germany). Five levels of scans were used to assess oocyte maturation (MII stage) and apoptosis. Because of incomplete penetration of the TUNEL stain (3–5 layers of cumulus cells), only the outer 2 layers of the cumulus complex were investigated regarding apoptosis. Apoptotic index was calculated as apoptotic cells/total cells visualised. Remaining oocytes were fertilized and cultured in vitro until Day 8. Day 7 and Day 8 blastocyst formation was assessed as well as blastocyst stage and grade. Effect of insulin treatment on variables was analysed by ANOVA following arc sin √p transformation. Post-ANOVA comparisons between H+L group v. Z were performed by using the contrast option under GLM (Scheffé test). Results are presented as least squares means ± s.e. P-values ≤ 0.05 were considered as statistically significant. Insulin treatment during oocyte maturation in vitro had no significant effect on oocyte nuclear maturation or apoptotic index of the cumulus cells (Z: 0.052 ± 0.025, L: 0.039 ± 0.016, H: 0.077 ± 0.044, P > 0.05). No effect was seen on cleavage rates (Z: 0.85 ± 0.02, L: 0.85 ± 0.02, H: 0.89 ± 0.03, P > 0.05), but insulin treatment significantly decreased Day 7 rates from fertilized oocytes (Z: 0.19 ± 0.02, L: 0.14 ± 0.02, H: 0.12 ± 0.02, P < 0.05). This study also showed a significantly retarded developmental stage and decreased grade of blastocysts in insulin-treated groups taken together when compared with the control group (P < 0.05). In this study, no effect of insulin supplementation during in vitro maturation was seen on bovine oocyte maturation and apoptosis of cumulus cells, but blastocyst formation and development were negatively affected. Further studies are needed for understanding the relationship between the addition of insulin during maturation in vitro and impaired blastocyst formation. Insulin is a common supplement in the first phase of the first in vitro maturation medium for pig oocytes and is believed to have a beneficial effect on this species.Funding was received from Stiftelsen Nils Lagerlöfs Fond H12–0051-NLA.

167 DEVELOPMENTAL COMPETENCE OF PORCINE OOCYTES DERIVED FROM SMALL- OR MEDIUM-SIZED FOLLICLES AND DENUDED OF CUMULUS CELLS BEFORE AND DURING IN VITRO MATURATION

2017 ◽  
Vol 29 (1) ◽  
pp. 192
Author(s):  
P. Ferré ◽  
K. X. Nguyen ◽  
T. Wakai ◽  
H. Funahashi
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Polar Body ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Cumulus Cells ◽  
Developmental Competence ◽  
Blastocyst Formation ◽  
First Polar Body

This experiment was undertaken to assess the meiotic and developmental competences of oocytes derived from different sized follicles and denuded of cumulus cells 0, 20, and 44 h after the start of culture for in vitro maturation (IVM). Groups of 60 oocyte-cumulus complexes from small- (SF; <3 mm) and medium-sized follicles (MF; 3–6 mm) were cultured for IVM in porcine oocyte medium with 50 μM β-mercaptoethanol supplemented with 1 mM dibutyryl-cyclic adenosine monophosphate, 10 IU mL−1 of eCG, and 10 IU mL−1 of hCG for 20 h at 39°C and 5% CO2 in air. Then, after washing, they continued culture in fresh β-mercaptoethanol without dibutyryl-cyclic adenosine monophosphate and gonadotropins under the same conditions for another 24 h. At 0, 20, and 44 h of IVM, cumulus cells were removed with 0.1% (wt/vol) hyaluronidase and the denuded oocytes continued IVM culture following the protocol. Mature oocytes with the first polar body were selected, parthenogenetically activated with a single electrical pulse (DC: 1.2 kV/cm, 30 µs), incubated with 4% (wt/vol) BSA and 5 μM cytochalasin B for 4 h, and cultured in porcine zygote medium for 5 days. Cleavage and blastocyst formation rates were observed on Day 2 and 5, respectively. Blastocysts were stained with 4’,6-diamidino-2-phenylindole for cell count assessment. The experiment was replicated 5 times and analysed with a 1- or 2-way ANOVA. If P < 0.05 in ANOVA, a Tukey multiple comparisons test was performed. Regardless of the time of cumulus cell removal, oocytes from MF had significantly higher in rates of maturation, cleavage, and blastocyst rates, as compared with those from SF, whereas there were no significant differences in the cell number of blastocysts between SF and MF (32 v. 34 cells, respectively). When oocytes were denuded before IVM culture, rates of oocyte maturation (37.6% in SF and 50.8% in MF), and blastocyst formation (2.7% in SF and 27.3% in MF) were significantly lower than controls (51.2% in SF and 76% in MF; 25.8% in SF and 48.5% in MF, respectively). When oocytes were denuded 20 h after the start of IVM, oocyte maturation rates were significantly increased (64.1% in SF and 82.5% in MF) as compared with controls, whereas no significant differences were observed in cleavage and blastocyst formation rates in comparison with controls. These results conclude that removing cumulus cells from oocyte-cumulus complexes 20 h after the start of IVM improves the meiotic competence of oocytes derived from both SF and MF, without any reduction of developmental competence of the oocytes following parthenogenetical activation.

166 MATURATION OF BOVINE OOCYTES IN POLY(DIMETHYLSILOXANE) MICROWELLS AND THEIR SUBSEQUENT DEVELOPMENT FOLLOWING IN VITRO FERTILIZATION

2014 ◽  
Vol 26 (1) ◽  
pp. 197
Author(s):  
K. Saeki ◽  
D. Iwamoto ◽  
S. Taniguchi ◽  
M. Kishi ◽  
N. Kato
Keyword(s):  
Oocyte Maturation ◽  
Subsequent Development ◽  
Cumulus Cells ◽  
Developmental Competence ◽  
High Humidity ◽  
Bovine Oocyte ◽  
Vitro Fertilization ◽  
Maturation Medium ◽  
Normal Fertilization

During bovine oocyte maturation, a lower density of cumulus cells surrounding oocytes reduces the developmental competence of the oocytes after IVF. Adding more cumulus cells (Hashimoto et al. 1998) rescues the developmental competence of the corona-enclosed oocytes. In this study, we examined the effects of poly(dimethylsiloxane) (PDMS) microwells (MW) for bovine oocyte maturation on the developmental competence of the oocytes following IVF. In experiment 1, MW were produced by making holes on 0.5-mm-thick PDMS plates using a 0.5-mm-diameter biopsy punch. The punched plates were placed on the bottoms of culture dishes. Bovine cumulus oocytes complexes (COC) were collected from slaughterhouse ovaries. Cumulus layers were removed from COC to prepare corona-enclosed oocytes (CEO) and denuded oocytes (DO). Then, COC, CEO, or DO were individually matured in single MW for 24 h at 39°C under 5% CO2 in air with high humidity. Ten oocytes of each group were matured in 50-μL droplets of maturation medium (group culture, GC) as controls. Maturation medium was TCM-199 supplemented with 10% FCS, 0.02 AU mL–1 FSH, and 1 μg mL–1 E2. The matured oocytes were fertilized with frozen–thawed spermatozoa. The embryos were cultured in CR1aa medium for 168 h under 5% CO2, 5% O2 and 90% N2 with high humidity. In experiment 2, effects of depth of MW for maturation on subsequent development following IVF were examined. Microwells were produced by making 0.5-mm-diameter holes on 0.5- or 1.5-mm-thick PDMS plates. Then, COC or CEO were individually matured in the MW for 24 h. Matured oocytes were fertilized in vitro and cultured for 168 h. Oocytes that were matured by GC were used as controls. In experiment 1(N = 4), rates of maturation (76–100%, n = 26 to 38), normal fertilization (53–70%, n = 44 to 49), and cleavage (61–77%, n = 114 to 117) were not different among all groups (P > 0.05; Fisher's PLSD test following ANOVA). Blastocyst rates were the same (P > 0.05) for COC matured in MW (50%) and by GC (43%). The rate for CEO that matured in MW (46%) tended to be higher (P = 0.061) than the rate for CEO that matured by GC (31%), and was comparable to the rate for COC matured by GC (43%). The blastocyst rates for DO that matured in MW and by GC were low (6%). In experiment 2 (N = 3), rates of maturation (86–100%, n = 13 to 28), normal fertilization (60–78%, n = 22 to 40), and cleavage (67–73%, n = 85 to 90) were not different among all groups (P > 0.05). However, the blastocyst rate for COC that matured in 1.5-mm-deep MW (53%) was significantly higher than the rates for COC that matured in 0.5-mm-deep MW (38%) and by GC (31%; P < 0.05). The results indicate that the developmental competence of oocytes that matured individually in PDMS MW was greater than that of oocytes that matured by GC. The deeper (1.5 mm) MW were found to be more effective for oocyte maturation than shallow (0.5 mm) MW and GC. The MW might increase density of cumulus cells surrounding oocytes, and the high cell-density enhanced the developmental competence of the oocytes.

O-062 In vitro maturation of oocytes in connection with ovarian tissue cryopreservation

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
M De Vos
Keyword(s):  
Cancer Patients ◽  
In Vitro Maturation ◽  
Ex Vivo ◽  
Ovarian Tissue ◽  
Ovarian Tissue Cryopreservation ◽  
Tissue Processing ◽  
Novel Approach ◽  
Tissue Cryopreservation

Abstract Abstract text Discussing fertility preservation (FP) in young cancer patients has become a key component of routine oncological health care. Although ovarian stimulation followed by oocyte cryopreservation has been recommended in cases where two to three weeks are available before the start of chemotherapy, ovarian tissue cryopreservation (OTC) is the preferred option when this timeframe is not available and when the potential gonadotoxic impact of cancer therapy is deemed moderate or severe, or in prepubertal girls. During ovarian tissue processing in the laboratory, cumulus-oocyte complexes can be identified. In vitro maturation and further vitrification of oocytes retrieved in ex vivo from the extracted ovarian tissue (ovarian tissue oocytes in vitro maturation; OTO-IVM) can be attempted to enhance the future reproductive options of the patient. Although the number of reported live births after OTO-IVM are limited, this experimental FP procedure has potential to become a standard appended procedure in conjunction with OTC. In cancer patients with haematological tumours and ovarian invasion, or patients with primary tumours of the ovary, ovarian tissue grafting may be contraindicated because of the risk of reintroducing malignant cells. Utilisation of vitrified oocytes after OTO-IVM may be the only hope for genetic offspring for these patients. Moreover, exogenous hormonal pretreatment is not required and COC can be recovered during ovarian tissue processing in the majority of patients who undergo partial or total unilateral oophorectomy. Nevertheless, maturation rates after OTO-IVM vary and are generally lower compared to IVM of transvaginally harvested IVM oocytes; currently available IVM systems registered for clinical use will have to be adapted to accommodate the in vitro requirements of oocytes derived from extracorporeal ovarian tissue, and follow-up data are needed to assess the success rate and safety of this novel approach.

scholarly journals The beneficial effects of cumulus cells and oocyte-cumulus cell gap junctions depends on oocyte maturation and fertilization methods in mice

PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e1761 ◽  
Author(s):  
Cheng-Jie Zhou ◽  
Sha-Na Wu ◽  
Jiang-Peng Shen ◽  
Dong-Hui Wang ◽  
Xiang-Wei Kong ◽  
...  
Keyword(s):  
Gap Junctions ◽  
Gap Junction ◽  
Oocyte Maturation ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Early Embryo ◽  
Blastocyst Formation ◽  
Beneficial Effects

Cumulus cells are a group of closely associated granulosa cells that surround and nourish oocytes. Previous studies have shown that cumulus cells contribute to oocyte maturation and fertilization through gap junction communication. However, it is not known how this gap junction signaling affectsin vivoversusin vitromaturation of oocytes, and their subsequent fertilization and embryonic development following insemination. Therefore, in our study, we performed mouse oocyte maturation and insemination usingin vivo- orin vitro-matured oocyte-cumulus complexes (OCCs, which retain gap junctions between the cumulus cells and the oocytes),in vitro-matured, denuded oocytes co-cultured with cumulus cells (DCs, which lack gap junctions between the cumulus cells and the oocytes), andin vitro-matured, denuded oocytes without cumulus cells (DOs). Using these models, we were able to analyze the effects of gap junction signaling on oocyte maturation, fertilization, and early embryo development. We found that gap junctions were necessary for bothin vivoandin vitrooocyte maturation. In addition, for oocytes maturedin vivo, the presence of cumulus cells during insemination improved fertilization and blastocyst formation, and this improvement was strengthened by gap junctions. Moreover, for oocytes maturedin vitro, the presence of cumulus cells during insemination improved fertilization, but not blastocyst formation, and this improvement was independent of gap junctions. Our results demonstrate, for the first time, that the beneficial effect of gap junction signaling from cumulus cells depends on oocyte maturation and fertilization methods.

scholarly journals The Differential Metabolomes in Cumulus and Mural Granulosa Cells from Human Preovulatory Follicles

2021 ◽  
Author(s):  
Er-Meng Gao ◽  
Bongkoch Turathum ◽  
Ling Wang ◽  
Di Zhang ◽  
Yu-Bing Liu ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Granulosa Cells ◽  
Quantitative Polymerase Chain Reaction ◽  
Cumulus Cells ◽  
Cholesterol Transport ◽  
Vitro Fertilization ◽  
Expression Of Genes ◽  
Preovulatory Follicles ◽  
Morphological Differences

AbstractThis study evaluated the differences in metabolites between cumulus cells (CCs) and mural granulosa cells (MGCs) from human preovulatory follicles to understand the mechanism of oocyte maturation involving CCs and MGCs. CCs and MGCs were collected from women who were undergoing in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) treatment. The differences in morphology were determined by immunofluorescence. The metabolomics of CCs and MGCs was measured by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) followed by quantitative polymerase chain reaction (qPCR) and western blot analysis to further confirm the genes and proteins involved in oocyte maturation. CCs and MGCs were cultured for 48 h in vitro, and the medium was collected for detection of hormone levels. There were minor morphological differences between CCs and MGCs. LC-MS/MS analysis showed that there were differences in 101 metabolites between CCs and MGCs: 7 metabolites were upregulated in CCs, and 94 metabolites were upregulated in MGCs. The metabolites related to cholesterol transport and estradiol production were enriched in CCs, while metabolites related to antiapoptosis were enriched in MGCs. The expression of genes and proteins involved in cholesterol transport (ABCA1, LDLR, and SCARB1) and estradiol production (SULT2B1 and CYP19A1) was significantly higher in CCs, and the expression of genes and proteins involved in antiapoptosis (CRLS1, LPCAT3, and PLA2G4A) was significantly higher in MGCs. The level of estrogen in CCs was significantly higher than that in MGCs, while the progesterone level showed no significant differences. There are differences between the metabolomes of CCs and MGCs. These differences may be involved in the regulation of oocyte maturation.

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