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1.
Thais S. Domingues Ana Paula Aquino Bruna Barros Raquel Mazetto Mariana Nicolielo Carolina M. Kimati Talita Devecchi Tatiana C. S. Bonetti Paulo C. Serafini Eduardo L. A. Motta 《Journal of assisted reproduction and genetics》2017,34(11):1553-1557
Purpose
Advances in reproductive techniques, mainly the introduction of oocyte vitrification, have provided the opportunity to conceive from oocyte banks. The aim of this study was to compare the clinical outcomes of fresh and vitrified oocytes in an egg donation program following blastocyst transfer.Methods
This retrospective observational study included 504 oocyte donation cycles. All donor women were younger than 30 years of age. The recipient cycles were divided into two groups: fresh oocytes (n = 78) or vitrified oocytes (n = 426). All oocytes were fertilized by ICSI using ejaculated sperm, followed by blastocyst transfer. Endometrium preparation was performed with estradiol valerate plus micronized progesterone according to standard protocols.Results
Recipients were of similar age (fresh 42.0 ± 4.5 years vs vitrified 41.8 ± 4.8 years; p = 0.790). The fresh group received more mature oocytes for injection compared to the vitrified group (10.1 ± 2.8 vs 9.2 ± 2.2; p = 0.005). The two pronuclei (2PN) rate (74.5 vs 77.4%; p = 0.195) and blastocyst rate (48.8 vs 51.6%; 0.329) were similar between the fresh and vitrified groups, respectively. The rates of clinical pregnancy were 60.9% in the fresh and 59.0% in the vitrified groups (p = 0.771).Conclusions
Our findings suggest that vitrified oocytes result in similar pregnancy rates when compared to fresh oocytes with blastocyst transfer in an egg donation program. Moreover, vitrified oocytes may allow for a better cycle schedule, starting with a lower number of oocytes to be fertilized. Therefore, we hypothesize that egg banks with vitrified oocytes could be safely utilized in an egg donation program.2.
The effects of platelet lysate on maturation,fertilization and embryo development of NMRI mouse oocytes at germinal vesicle stage 下载免费PDF全文
Hassan Pazoki Hussein Eimani Farah Farokhi Abdol-Hossein Shahverdi Leila Sadat Tahaei 《Reproductive Medicine and Biology》2016,15(2):115-120
Purpose
Improving in vitro maturation could increase the rate of pregnancy from oocytes matured in vitro. Consequently, patients will be prevented from using gonadotropin with its related side effects. In this study, the maturation medium was enriched by platelet lysate (PL), then maturation and subsequent developments were monitored.Methods
Oocytes at germinal vesicle stage with cumulus cells (cumulus–oocyte complex) and without cumulus cells (denuded oocytes) were obtained from mature female mice. The maturation medium was enriched by 5 and 10 % PL and 5 % PL + 5 % fetal bovine serum (FBS) as experimental groups; the control groups’ media consisted of 5 and 10 % FBS. After 18 h, the matured oocytes were collected and, after fertilization, subsequent development was monitored.Results
The rates of maturation, fertilization and 2-cell embryo development for the denuded oocyte groups in experimental media 5 % PL and 5 % PL + 5 % FBS were significantly higher than those of the control groups (P < 0.05), while the results for the cumulus–oocyte complex groups were similar between the experimental groups and control groups.Conclusions
The results of this study indicated that platelet lysate could improve the maturation rate in the absence of granulosa cells compared to media with FBS. This extract also had positive effects on fertilization and embryo development.3.
Background
Metformin decreases polycystic ovary syndrome (PCOS) symptoms, induces ovulation, and may improve developmental competence of in vitro oocyte maturation. This study was designed to define the effects of metformin on the characteristics of in vitro oocyte maturation in estradiol valerate (EV) PCOS-induced rats.Methods
Forty-five adult female Sprague–Dawley rats were randomly divided into control; sham and PCOS-induced (treated by a single dose of estradiol valerate, 4 mg/rat, IM) groups. The body weight was measured weekly for 12 weeks. At the end of week 12, the serum levels of testosterone, estrogen, progesterone, LH, and FSH and blood glucose of all the rats were measured. About 380 cumulus oocyte complexes (control, 125; sham, 122; PCOS-induced rats, 133) were incubated in Ham’s F10 in the absence and/or presence of metformin (M 5?10) for 12, 24, 36, and 48 h. The cumulus cells expansion and nuclear and cytoplasmic maturation of the oocytes was evaluated using 1 % aceto-orcein staining, and transmission electron microscopy (TEM).Results
No significant differences were observed in the body weight of the rats. The serum level of testosterone was reduced, and progesterone and LH were significantly increased in the PCOS-induced rats (p?<?0.05). However, no significant differences were observed in the serum levels of estrogen and FSH among the groups. Blood glucose level was higher in the PCOS-induced rats than control, (p?<?0.01). The expansion of cumulus cells was observed in the metformin-treated oocytes. The oocytes retrieved from PCOS-induced rats show a stage of meiotic division (GVBD, MI, A-T, and MII) in 57.12 % of metformin-untreated and fairly significantly increased to 64.28 % in metformin-treated oocytes, (p?<?0.05), but no differences were observed in the MII stage within groups. The redistribution of some cytoplasmic organelles throughout the ooplasm, particularly the peripheral cortical granules, was defined in the metformin-treated oocytes.Conclusions
Single dose of EV can creates a reversible PCO adult rat model. Metformin enhances the COCs to initiate meiotic resumption at the first 6 h of IVM. In our study the metformin inability to show all aspects of in vitro oocyte maturation and may be resulted from deficiency of EV to induce PCOS.4.
Arindam Dhali Pradeep Krishna Javvaji Atul P. Kolte Joseph Rabinson Francis Sudhir C. Roy Veerasamy Sejian 《Journal of assisted reproduction and genetics》2017,34(11):1493-1500
Purpose
Cumulus cells (CC) play important roles in oocyte development and cumulus expressed genes can be used as markers for oocyte quality. This study aimed to investigate temporal changes in the expression of cumulus marker genes during oocyte maturation as possible biomarkers of embryo developmental competence in ovine.Methods
Gene expression was assessed in the CC of the BCB+ (developmentally competent) and BCB- (developmentally poor) oocytes at 0, 12, and 24 h of in vitro maturation (IVM). Further, the association between the temporal cumulus gene expression and in vitro oocyte and embryo development was assessed.Results
The maturation and blastocyst formation rates were found significantly greater for the BCB+ than the BCB- oocytes. At the 0 h of IVM, a significant upregulation in the expression of PTGS2, STAR, SDC2, LHR, FGF2, BCL2, IL7RA, HSPA1A, and IFNT was observed in the CC of the poor (BCB-) as compared to the competent (BCB+) oocytes. In contrast, it was observed that as maturation progressed, the cumulus expression of most of the favorable genes was reduced and was found significantly downregulated at the completion of IVM in the poor as compared to the competent oocytes.Conclusions
The study revealed noticeable differences in the cumulus gene expression profile at different stages of IVM between ovine oocytes of differential developmental ability. The results indicated that the loss of cumulus gene expression along the maturation period in the poor oocytes was related to their intrinsic poor quality in the ovarian follicle.5.
Huiqun Yin Hong Jiang Stine Gry Kristensen Claus Yding Andersen 《Journal of assisted reproduction and genetics》2016,33(6):741-746
Purpose
The aim of the study was to investigate the maturation rate of immature oocytes collected from ovarian medulla tissue normally discarded during preparation of ovarian cortical tissue for fertility preservation. Further we evaluated survival of derived MII oocytes following vitrification and warming.Methods
36 patients aged from 8 to 41 years who had one ovary excised for fertility preservation were included. Oocytes were collected from the medulla tissue and matured in vitro 44–48 h followed by vitrification. Number of oocytes collected, the rates of maturation and post-warming survival were assessed.Results
On average, 11 immature oocytes were collected per patient. The overall maturation rate was 29 % irrespective of whether the ovary was transported 4–5 h on ice or obtained immediately after oophorectomy. The maturation rate in patients below 20 years of age (55 %) was significantly higher than that of patients aged 20–30 years (29 %) and above 30 years (26 %). The post-warming survival rate was 64 %. No significant relationship was observed between the number of collected oocytes and the age of patients.Conclusions
Approximately three MII oocytes were obtained per patient following in vitro maturation (IVM) of immature oocytes collected from medulla tissue, of which two survived vitrification and warming. This approach represents an add-on method to potentially augment the fertility opportunity for cancer patients, especially in young women with cancer where transplantation of cortical tissue may pose a risk of relapse, but the IVM approach is currently too inefficient to be the only method used for fertility preservation.6.
7.
8.
Yoni Cohen Alexandra St-Onge-St-Hilaire Samer Tannus Grace Younes Michael H. Dahan William Buckett Weon-Young Son 《Journal of assisted reproduction and genetics》2018,35(9):1683-1689
Purpose
To assess effects on fertilization rate, embryo quality, pregnancy, and live birth rates of vitrification and warming of oocytes that matured in vitro (vIVM) compared to fresh in vitro maturation (fIVM) cycles.Methods
A retrospective cohort study conducted at a university hospital-affiliated IVF unit. Fifty-six cycles of vIVM cycles and 263 fIVM in women diagnosed with polycystic ovarian syndrome (PCOS) ovaries were included in the analysis. The study group included PCOS patients who failed ovulation induction with intrauterine insemination and were offered IVM cycle followed by oocyte vitrification and warming. The embryological aspects and clinical outcomes were compared to those of controls undergoing fresh IVM cycles during the same period. The main outcome measure was live birth rate.Results
One thousand seventy oocytes were collected from 56 patients and underwent vitrification and warming. In the control group, 4781 oocytes were collected from 219 patients who had undergone a fresh IVM cycle. Oocyte maturation rates were similar between the groups (mean ± SD: 0.7?±?0.2 vs. 0.6?±?0.2, for vIVM and fIVM, respectively). Survival rate after warming was 59.8%. Fertilization and embryo cleavage rates per oocyte were significantly lower in the vIVM group. Clinical pregnancy (10.7 vs. 36.1%) and live birth rates (8.9 vs. 25.9%) per cycle were significantly lower in the vIVM group than those in the fIVM group (P?=?0.005 and P?<?0.001, respectively). Five healthy babies were born in the vIVM group.Conclusions
The reproductive potential of vitrified IVM oocytes is impaired. This injury likely occurs through vitrification and warming.9.
10.
I. A. P. Derks-Smeets T. C. van Tilborg A. van Montfoort L. Smits H. L. Torrance M. Meijer-Hoogeveen F. Broekmans J. C. F. M. Dreesen A. D. C. Paulussen V. C. G. Tjan-Heijnen I. Homminga M. M. J. van den Berg M. G. E. M. Ausems M. de Rycke C. E. M. de Die-Smulders W. Verpoest R. van Golde 《Journal of assisted reproduction and genetics》2017,34(11):1475-1482
Purpose
The aim of this study was to determine whether BRCA1/2 mutation carriers produce fewer mature oocytes after ovarian stimulation for in vitro fertilization (IVF) with preimplantation genetic diagnosis (PGD), in comparison to a PGD control group.Methods
A retrospective, international, multicenter cohort study was performed on data of first PGD cycles performed between January 2006 and September 2015. Data were extracted from medical files. The study was performed in one PGD center and three affiliated IVF centers in the Netherlands and one PGD center in Belgium. Exposed couples underwent PGD because of a pathogenic BRCA1/2 mutation, controls for other monogenic conditions. Only couples treated in a long gonadotropin-releasing hormone (GnRH) agonist-suppressive protocol, stimulated with at least 150 IU follicle stimulating hormone (FSH), were included. Women suspected to have a diminished ovarian reserve status due to chemotherapy, auto-immune disorders, or genetic conditions (other than BRCA1/2 mutations) were excluded. A total of 106 BRCA1/2 mutation carriers underwent PGD in this period, of which 43 (20 BRCA1 and 23 BRCA2 mutation carriers) met the inclusion criteria. They were compared to 174 controls selected by frequency matching.Results
Thirty-eight BRCA1/2 mutation carriers (18 BRCA1 and 20 BRCA2 mutation carriers) and 154 controls proceeded to oocyte pickup. The median number of mature oocytes was 7.0 (interquartile range (IQR) 4.0–9.0) in the BRCA group as a whole, 6.5 (IQR 4.0–8.0) in BRCA1 mutation carriers, 7.5 (IQR 5.5–9.0) in BRCA2 mutation carriers, and 8.0 (IQR 6.0–11.0) in controls. Multiple linear regression analysis with the number of mature oocytes as a dependent variable and adjustment for treatment center, female age, female body mass index (BMI), type of gonadotropin used, and the total dose of gonadotropins administered revealed a significantly lower yield of mature oocytes in the BRCA group as compared to controls (p = 0.04). This finding could be fully accounted for by the BRCA1 subgroup (BRCA1 mutation carriers versus controls p = 0.02, BRCA2 mutation carriers versus controls p = 0.50).Conclusions
Ovarian response to stimulation, expressed as the number of mature oocytes, was reduced in BRCA1 but not in BRCA2 mutation carriers. Although oocyte yield was in correspondence to a normal response in all subgroups, this finding points to a possible negative influence of the BRCA1 gene on ovarian reserve.11.
Ioannis A. Sfontouris Efstratios M. Kolibianakis George T. Lainas Christos A. Venetis George K. Petsas Basil C. Tarlatzis Tryfon G. Lainas 《Journal of assisted reproduction and genetics》2017,34(10):1377-1383
Purpose
The aim of this study is to determine whether blastocyst utilization rates are different after continuous culture in two different commercial single-step media.Methods
This is a paired randomized controlled trial with sibling oocytes conducted in infertility patients, aged ≤40 years with ≥10 oocytes retrieved assigned to blastocyst culture and transfer. Retrieved oocytes were randomly allocated to continuous culture in either Sage one-step medium (Origio) or Continuous Single Culture (CSC) medium (Irvine Scientific) without medium renewal up to day 5 post oocyte retrieval. Main outcome measure was the proportion of embryos suitable for clinical use (utilization rate).Results
A total of 502 oocytes from 33 women were randomly allocated to continuous culture in either Sage one-step medium (n = 250) or CSC medium (n = 252). Fertilization was performed by either in vitro fertilization or intracytoplasmic sperm injection, and embryo transfers were performed on day 5. Two patients had all blastocysts frozen due to the occurrence of severe ovarian hyperstimulation syndrome. Fertilization and cleavage rates, as well as embryo quality on day 3, were similar in the two media. Blastocyst utilization rates (%, 95% CI) [55.4% (46.4–64.1) vs 54.7% (44.9–64.6), p = 0.717], blastocyst formation rates [53.6% (44.6–62.5) vs 51.9 (42.2–61.6), p = 0.755], and proportion of good quality blastocysts [36.8% (28.1–45.4) vs 36.1% (27.2–45.0), p = 0.850] were similar in Sage one-step and CSC media, respectively.Conclusions
Continuous culture of embryos in Sage one-step and CSC media is associated with similar blastocyst development and utilization rates. Both single-step media appear to provide adequate support during in vitro preimplantation embryo development. Whether these observations are also valid for other continuous single medium protocols remains to be determined.Clinical trial registration number: NCT02302638.12.
Yuval Atzmon Ester Shoshan-Karchovsky Medeia Michaeli Nardin Aslih Guy Shrem Adrian Ellenbogen Einat Shalom-Paz 《Journal of assisted reproduction and genetics》2017,34(9):1145-1151
Background
Obesity is associated with several fertility disorders. This prospective cohort study was designed to evaluate the effect of body mass index (BMI) (kg/m2) on oocyte diameter and treatment.Methods
Women undergoing in vitro fertilization (IVF) with intracytoplasmic sperm injection (ICSI) were enrolled in the study. They were divided into two groups according to BMI: obese (BMI > 30) and normal weight (BMI < 25). Mature oocytes were evaluated according to total diameter, zona pellucida, and oolema diameters.Results
A total of 387 oocytes were obtained from the 46 women who participated. Significantly more mature oocytes (M2) were retrieved from normal weight patients compare to obese women (15.1 ± 6.8 vs. 9.7 ± 3.9, respectively, P < 0.001). Oocytes from women in the obese group were significantly smaller than those in the normal weight group, including oocyte diameter (157.9 ± 7.9 vs. 164.3 ± 5.1 μm, P < 0.0001), oolema diameter (110.3 ± 4.5 vs. 113.5 ± 3.5 μm, P < 0.0001), and zona pellucida thickness (17.9 ± 2.6 vs. 19.0 ± 2.4 μm, P < 0.000), respectively. Multivariate logistic regression analysis, including oolema diameter, female age, BMI, number of M2 oocytes, and zona pellucida, was conducted to predict pregnancy. Small oolema diameter in obese patient adversely correlated with pregnancy. Larger oolema diameter was positively associated with the probability of pregnancy in the obese group as well as thinner zona pellucida.Conclusion
Obesity is associated with smaller oocytes, which adversely affect fertility outcomes.Trial registration
NIH number NCT0167293113.
Debora A. Anzalone Domenico Iuso Marta Czernik Grazyna Ptak Pasqualino Loi 《Journal of assisted reproduction and genetics》2016,33(6):757-763
Purpose
This study aims to determine if the integrity of the sperm plasma membrane and acrosome vesicle could be limiting factors in sheep intracytoplasmic sperm injection (ICSI).Methods
Prior to in vitro fertilization (IVF) or ICSI, the oocytes were subjected to in vitro maturation (IVM) for 24 h. First, to evaluate the need of artificial activation for ovine ICSI, 226 oocytes were injected with intact spermatozoa (IS), from which 125 were activated by incubation in ionomycin and 101 were cultured without activation. Next, spermatozoa were mechanically (by piezo-electrical pulses) and/or chemically (by ionomycin/Triton X-100) treated to break membranes and acrosomes and were injected into oocytes, grouped as follows: (i) piezo-pulsed spermatozoa (PPS), (ii) PPS pre-treated with ionomycin (PPS-I), (iii) PPS pre-treated with Triton X-100 (PPS-T), and (iv) intact and untreated spermatozoa as a control (CTR-IS).Results
No differences were observed in the zygote/cleavage/blastocyst rate between chemically activated and non-activated oocytes (50 vs. 45 %, 11.6 vs. 10.1 %; 1.8 vs. 1.1 %, respectively), after ICSI with CTR-IS. Injection of PPS compared to CTR-IS increased the proportion of zygotes and blastocysts (84.6 vs. 45 %, p?<?0.01; 15.5 vs. 1.1 %, p?<?0.0001, respectively). Moreover, the percentage of PPS-derived blastocysts was not significantly different from that obtained by conventional IVF (15.5 vs. 20.2 %). The ICSI blastocysts’ development was also improved with PPS pre-treated with ionomycin (15.6 %), but was completely impeded with PPS pre-treated with Triton X-100 (0 %).Conclusion
Our findings confirm that ICSI with spermatozoa whose plasma membrane and acrosome have been mechanically damaged substantially improves embryonic development until the blastocyst stage.14.
Lucie Rolland Jeanne Perrin Virginie Villes Valérie Pellegrin Léon Boubli Blandine Courbiere 《Journal of assisted reproduction and genetics》2017,34(11):1523-1528
Purpose
Does the type of anesthesia (paracervical block (PCB) or general anesthesia (GA)) impact live birth rate, pain, and patient satisfaction?Methods
A non-randomized prospective cohort study was conducted in women treated for IVF. Two groups of patients were prospectively included: the PCB group (n = 234) and the GA group (n = 247). The type of anesthesia was determined by the patients. The primary endpoint was cumulative live birth rate by OR. Secondary endpoints were self-assessment of the patients’ peri-operative abdominal and vaginal pain vs the doctors’ evaluations during PCB, post-operative abdominal and vaginal pain level, and patient satisfaction in both groups. Pain levels were assessed with a numerical rating scale (NRS).Results
The live birth rate was similar in both groups (19.8% in the GA group vs 20.9% in the PCB group, P = 0.764). During oocyte retrieval in the PCB group, the physicians significantly under-estimated the vaginal pain experienced by the patients (3.04 ± 0.173 for patients vs 2.59 ± 0.113 for surgeons, P = 0.014). Post-operative vaginal and abdominal pain were significantly greater in the PCB group compared to the GA group (2.26 ± 0.159 vs 1.66 ± 0.123, respectively, P = 0.005, and 3.80 ± 0.165 vs 3.00 ± 0.148, respectively, P < 0.001). Patients were more significantly satisfied with GA than with PBC (P < 0.001).Conclusion
Because the LBR was similar in both groups and patient satisfaction was high, the choice of anesthesia should be decided by the patients.15.
Farideh Yazdanpanah Mohammad Ali Khalili Maryam Eftekhar Hojatallah Karimi 《Archives of gynecology and obstetrics》2013,288(2):439-444
Background
15 % of oocytes collected from Assisted Reproductive Technology (ART) cycles are immature. These oocytes may be matured following in vitro maturation (IVM) program. It is possible to cryopreserve the immature oocytes for further use in ART after application of IVM.Objective
The aim was to determine the maturation rate and viability of human oocytes that were matured in vitro after vitrification program.Materials and methods
63 women (19–43 years old) who underwent controlled ovarian stimulation for ART were included in this study. 53 immature oocytes were used for fresh group (fIVM) and 50 immature oocytes for vitrification group (vIVM). The maturation medium was Ham’s F10 supplemented with 0.75 IU FSH, 0.75 IU LH and 40 % human follicular fluid (HFF). After 36 h, maturation and morphology of all oocytes were assessed. Also, the oocyte viability was assessed using PI/Hoechst immunostaining technique.Results
The maturation rates were reduced in vIVM group (56.0 %) in comparison to fIVM group (88.7 %; P < 0.001). Oocyte viability rate were also reduced in vIVM group (56.0 %) in comparison to fIVM (86.8 %, P < 0.007).Conclusions
Cryopreservation via vitrification reduced both the maturation capacity and viability of human oocytes in IVM technology. It is, therefore, recommended to apply IVM on fresh immature oocytes, instead. 相似文献16.
Przemysław Ciepiela Antoni J. Dulęba Ewelina Kowaleczko Kornel Chełstowski Rafał Kurzawa 《Journal of assisted reproduction and genetics》2018,35(7):1265-1276
Purpose
This study investigated the relationship between the vitamin D [25(OH)D] level in individual follicles and oocyte developmental competence.Methods
A prospective cohort study in a private infertility center. Infertile women (N?=?198) scheduled for intracytoplasmic sperm injection (ICSI) and a single embryo transfer (SET) provided serum samples and 322 follicular fluid (FF) specimens, each from a single follicle on the day of oocyte retrieval.Results
FFs corresponding to successfully fertilized oocytes (following ICSI) contained significantly lower 25(OH)D level compared with those that were not fertilized (28.4 vs. 34.0 ng/ml, P?=?0.001). Top quality embryos on the third day after fertilization, when compared to other available embryos, developed from oocytes collected from follicles containing significantly lower 25(OH)D levels (24.56 vs. 29.59 ng/ml, P?=?0.007). Positive hCG, clinical pregnancy, and live birth rates were achieved from embryos derived from oocytes that grew in FF with significantly lower 25(OH)D levels than in follicles not associated with subsequent pregnancy. The concentration of 25(OH)D in FF in women with negative hCG was 32.23?±?20.21 ng/ml, positive hCG 23.62?±?6.09 ng/ml, clinical pregnancy 23.13?±?6.09 ng/ml, and live birth 23.45?±?6.11 ng/ml (P?<?0.001). Women with serum 25(OH)D?<?20 ng/ml had not only a higher fertilization rate (71 vs. 61.6%, P?=?0.026) and a higher clinical pregnancy rate (48.2 vs. 25%, P?=?0.001), but also higher miscarriage rate (14.5 vs. 3.8%, P?=?0.013) compared with those with levels ≥?20 ng/ml.Conclusion
This study reveals that the level of 25(OH)D in FF correlates negatively with the oocytes’ ability to undergo fertilization and subsequent preimplantation embryo development. Oocytes matured in FF with low 25(OH)D concentration are more likely to produce top quality embryos and are associated with higher pregnancy and delivery rates. On the other hand, low serum vitamin D concentration is associated with higher miscarriage rates.17.
Xin Tao Jessica N. Landis Rebecca L. Krisher Francesca E. Duncan Elena Silva Agnieszka Lonczak Richard T. ScottIII Yiping Zhan Tinchun Chu Richard T. ScottJr Nathan R. Treff 《Journal of assisted reproduction and genetics》2017,34(12):1587-1594
Purpose
It was reported that mitochondrial DNA (mtDNA) was significantly increased in aneuploid human embryos compared to euploid embryos and was also associated with maternal age. In this study, we further established the mouse model of mtDNA quantitation in reproductive samples based on whole-genome amplification (WGA) and next-generation sequencing (NGS).Methods
WGA followed by NGS-based mtDNA quantitation was first performed on 6 single- and 100-cell samples from a tumor-derived mouse cell line, which was exposed to ethidium bromide to reduce mtDNA content. The relative mtDNA content was normalized to nuclear DNA. This method was then applied to mouse reproductive samples, including 40 pairs of oocytes and polar bodies from 8 CD-1 female mice of advanced reproductive age and 171 blastocysts derived via in vitro maturation (IVM) or in vivo maturation (IVO) from young (6–9 weeks) and reproductively aged (13.5 months) female CF-1 mice.Results
Exposure to ethidium bromide for 3 and 6 days decreased mtDNA levels in both the single- and 100-cell samples as expected. Results demonstrated that the first polar body contained an average of 0.9% of mtDNA relative to oocytes. Compared to the cells in blastocysts, oocytes contained about 180 times as much mtDNA per cell. mtDNA levels were compared among blastocysts from reproductively young and old female mice that had either been produced by IVM or IVO. Cells in blastocysts from younger mice contained significantly lower amounts of mtDNA compared to aged mice (P < 0.0001). Cells in blastocysts produced via IVO had higher mtDNA content than IVM-derived blastocysts (P = 0.0001). Cells in aneuploid blastocysts were found to have significantly higher (1.74-fold) levels of mtDNA compared to euploid blastocysts (P = 0.0006).Conclusion
A reliable method for assessing mtDNA content in mouse gametes and embryos was established. Relative mtDNA levels were elevated in aneuploid embryos relative to euploid embryos, were higher in blastocysts from reproductively old mice relative to young mice, and were lower in embryos derived from IVM compared to IVO.18.
Yamato Mizobe Naoto Oya Reiko Iwakiri Naomi Yoshida Yumi Sato Nanase Onoue Kazuchika Miyoshi Makoto Tokunaga Yuji Ezono 《Journal of assisted reproduction and genetics》2016,33(12):1685-1690
Purpose
In intracytoplasmic sperm injection (ICSI) of oocytes with a fragile oolemma (fragile oocytes), breakage can occur at injection. In this study, we produced embryos from oocytes with a fragile and normal oolemma (normal oocytes) by ICSI and compared their ability to be fertilized and develop in vitro. We also investigated whether fragile oocyte-derived embryos could implant after blastocyst transfer to determine whether fragile oocytes should be used for assisted reproductive technology treatment.Methods
Oocytes were divided into three groups—normal oocytes from cycles containing no fragile oocytes (group A), normal oocytes from cycles containing at least one fragile oocyte (group B), and fragile oocytes (group C), and their fertilization abilities after ICSI and the developmental abilities of resultant embryos were compared.Results
The fertilization rate in group C (65.3 %) was significantly (P?<?0.01) lower than those in groups A (84.6 %) and B (86.9 %), and the degeneration rate in group C (24.2 %) was significantly (P?<?0.01) higher than those in groups A (0.71 %) and B (0.28 %). However, there were no significant differences in the blastocyst formation rates (59.7–67.5 %) of embryos among the different groups. In addition, the pregnancy rate after transfer of blastocysts in group C (50.0 %) was not significantly different from those in groups A (35.6 %) and B (45.8 %).Conclusions
The fertilization ability after ICSI of fragile oocytes is lower than that of normal oocytes but the resultant embryos have the same developmental ability as those of normal oocyte-derived embryos.19.
Yijuan Sun Ruihuan Gu Xiaowei Lu Shen Zhao Yun Feng 《Journal of assisted reproduction and genetics》2016,33(2):231-236
Purpose
The aim of this study is to evaluate the impact of oocyte vitrification on embryo development potential and to assess the chromosome abnormalities of blastocysts derived from fresh/vitrified-warmed oocytes to assure the safety of the oocyte cryopreservation technique.Methods
In vitro matured oocytes derived from immature oocytes were retrieved from small follicles during IVF/intracytoplasmic sperm injection (ICSI) cycles were randomly divided into a fresh and vitrified-warmed groups. After intracytoplasmic sperm injection, the fertilization rate, embryo quality, and developmental status were compared between the two groups. Blastocysts derived from both groups were analyzed using the copy number variation (CNV)-seq technique to evaluate DNA abnormalities.Results
The fertilization rate with ICSI and the cleavage rate were similar between the two groups. Among the vitrified-warmed group, there was a lower incidence of usable embryos on day 3 (16.42 vs. 28.57 %; P?<?0.05) and a lower incidence of blastocysts (7.46 vs. 17.86 %; P?<?0.05). However, the proportions of embryos that developed to blastocysts from the day 3 available embryos were similar between the two groups (62.5 vs. 45.45 %; P?>?0.05). In the day 3 embryos, the proportion of >5 cell embryos in the fresh group was markedly higher than in the vitrified-warmed group (41.67 vs. 21.64 %; P?<?0.05), and the proportion of embryos with ≧50 % fragments was not significantly different between the two groups (39.29 vs. 43.28 %; P?>?0.05). The result of CNV-seq demonstrated that there was no difference in chromosomal abnormalities between the two groups (20 vs. 20 %).Conclusions
Oocyte vitrification and the warming procedure diminished the embryo development potential before day 3, when embryo genomic activation started. The day 3 usable embryos derived from vitrified-warmed oocytes had the same potential for developing into blastocysts. Vitrification and the warming procedure did not increase the chromosome abnormalities of the blastocysts. Oocyte vitrification is a safe technique for those patients who have no other options, although the oocyte efficiency may be diminished after the vitrified-warmed procedure.20.
Thi Ngoc Lan Vuong M. T. Ho T. Q. Ha M. Brehm Jensen C. Yding Andersen P. Humaidan 《Journal of assisted reproduction and genetics》2017,34(4):471-478