scholarly journals Sucrose ‘Versus’ Trehalose Cryoprotectant Modification in Oocyte Vitrification : A Study of Embryo Development

2018 ◽  
Vol 11 (1) ◽  
pp. 97-104 ◽  
Author(s):  
Silvia W. Lestari ◽  
Khairunnisa F. Ilato ◽  
M. Iqbal A. Pratama ◽  
Nurin N. Fitriyah ◽  
Mulyoto Pangestu ◽  
...  

Numerous studies reported that vitrification, an ultra-rapid cooling technique, seems to be highly effective and could increase oocyte survival rate rather than slow freezing. The successful of oocyte vitrification depends on the proper combination of type and concentration of cryoprotectant. This study was addressed to determine the effects of the combination of type and concentration of cryoprotectants of vitrification media, notably in the embryo development. This experimental research was conducted by using oocyte obtained from thirty-two adult female Deutschland, Denken and Yoken (DDY) mice (7-8 weeks old). The MII mice oocytes were vitrified within 24 h after retrieval using the Cryotop method with cryoprotectants as follow : sucrose (16.5% EG, 16.5% DMSO, 0.5 mol/l sucrose), trehalose (16.5% EG, 16.5% DMSO, 0.5 mol/l trehalose) and Kitazato. The embryo development and morphological grading was observed at 2-cell and 8-cells under reverse phase light microscope and inverted microscope. This study demonstrated a good embryo development and morphological grading in sucrose and trehalose vitrification media. In embryo development, trehalose medium seems more superior compared to sucrose medium, even though Kitazato was the most superior compared to both. In the morphological grading, in 2-cells embryo, there were no significant differences between the three cryoprotectants, While, in 8-cells embryo, trehalose medium appeared to be superior compared to sucrose medium, even though seemed more inferior compared to Kitazato. The appropriate type and concentration of sugar as extracellular cryoprotectant was trehalose in oocyte vitrification based on embryo development, compared to sucrose.

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
S Montgomery ◽  
K Montgomery ◽  
D Nash ◽  
A Campbell

Abstract Study question Are the morphokinetic profiles, as assessed using time-lapse technology, of human embryos developed from vitrified oocytes different to those from fresh oocytes. Summary answer Vitrification of oocytes does have an effect on early developmental morphokinetic profiles, but this is normalized by the time the embryo has reached blastocyst. What is known already Vitrification of oocytes is now commonplace, but little is known about the effect this may have on subsequent embryo development. Study design, size, duration This was a retrospective data analysis, from 8 fertility clinics in the UK between 2012 and 2019. Embryos from patients in the vitrified group (n = 557) were matched to fresh patient controls (n = 539). The matching was performed based on the following criteria: type of treatment, patient age, cause of infertility and number of embryos. Participants/materials, setting, methods The embryos in each group were compared for mean morphokinetics of key developmental stages in hours post insemination (hpi). Parameters compared included early cleavage divisions (t2-t8), time to start of compaction (tSC), time to morula (tM), time to start blastulation (tSB), time to full blastocyst (tB) and duration of compaction (tB-tSC). Treatment outcomes were compared between the two groups, including percentage of blastocyst formation, clinical pregnancy rate, implantation rate and live birth rate. Main results and the role of chance The results showed a significant delay across all early cleavage divisions as follows for vitrified and fresh oocytes respectively: 2-cell (28.14 vs 26.10 (p < 0.001)), 3 cell (37.56 vs 35.37 (p < 0.001)), 4 cell (40.58 vs 37.54 (p < 0.001)), 5 cell (50.31 vs 47.14 (p < 0.001)), 6 cell (53.99 vs 50.87 (p < 0.001)), 7 cell (57.08 vs 54.48 (p < 0.001)) and 8 cell (61.26 vs 58.91 (p < 0.01)). In addition, tSC was also significantly delayed in the vitrified group (80.65 vs 76.36 (p < 0.001)). However, the compaction stage was significantly shorter in the vitrified oocytes (19.02 vs 22.45 (p < 0.001)). Therefore, there was no difference in the time that embryos derived from fresh and vitrified oocytes reached the blastocyst stage (108.03 vs 107.78 (p > 0.05)). No difference was found in clinical pregnancy, implantation or live birth rates but significantly fewer blastocyst developed from vitrified oocytes compared to fresh (36.09% vs 42.4% (p < 0.05)). Limitations, reasons for caution Although this was a matched analysis, it was a retrospective in nature therefore is subject to confounders. However, it would be problematic to perform a prospective randomized controlled trial to address this study question given the need to randomize patients to elective freezing of oocytes prior to embryo creation. Wider implications of the findings: Vitrification of oocytes may affect early developmental morphokinetic profiles, but any effect is normalized by the time the embryo has reached blastocyst. However, fewer blastocysts may develop following oocyte vitrification. This may have implications for oocyte donation banks and those patients choosing to cryopreserve oocytes. Trial registration number NA


Author(s):  
ADITYA SANKAR

Abstract This experiment describes the in vitro transcription of Kdm4a wildtype and H188A catalytic dead mRNA. This details also its subsequent injection into mouse oocytes followed my IVF to track the impact on embryo development. The procedure is technically challenging and performed by the Transgenic Core Facility at the University of Copenhagen. Oocytes have a poorer survival rate following mRNA inject as against zygotes. However the objective was to demonstrate the earliest stage of intervention to rescue developmental failure of KDM4A maternal zygotic mutant embryos


2020 ◽  
Vol 55 (10) ◽  
pp. 1328-1336
Author(s):  
Valentina I. Mokrousova ◽  
Konstantin A. Okotrub ◽  
Eugeny Y. Brusentsev ◽  
Elena A. Kizilova ◽  
Nikolai V. Surovtsev ◽  
...  

Reproduction ◽  
2006 ◽  
Vol 131 (4) ◽  
pp. 795-804 ◽  
Author(s):  
Wen-Qing Shi ◽  
Shi-En Zhu ◽  
Dong Zhang ◽  
Wei-Hua Wang ◽  
Guo-Liang Tang ◽  
...  

This study was designed to examine the effect of Taxol pretreatment on vitrification of porcine oocytes matured in vitro by an open pulled straw (OPS) method. In the first experiment, the effect of Taxol pretreatment and fluorescein diacetate (FDA) staining on parthenogenetic development of oocytes was evaluated. In the second experiment, viability, microtubule organization and embryo development of oocytes were assessed after oocytes were exposed to vitrification/warming solutions or after vitrification with or without Taxol pretreatment. The results showed that Taxol pretreatment and/or FDA staining did not negatively influence the oocyte’s developmental competence after parthenogenetic activation. After being exposed to vitrification/warming solutions, the survival rate (83.3%) of the oocytes was significantly (P < 0.05) reduced as compared with that in the control (100%). Vitrification/warming procedures further reduced the survival rates of oocytes regardless of oocytes being treated with (62.1%) or without (53.8%) Taxol. The proportions of oocytes with normal spindle configuration were significantly reduced after the oocytes were exposed to vitrification/warming solutions (38.5%) or after vitrification with (10.3%) or without (4.1%) Taxol pretreatment as compared with that in control (76.8%). The rates of two-cell-stage (5.6–53.2%) embryos at 48 h and blastocysts (0–3.8%) at 144 h after activation were significantly reduced after exposure to vitrification/warming solutions or after vitrification as compared with control (90.9% and 26.6% respectively). However, the proportion of vitrified oocytes developed to two-cell stage was significantly higher when oocytes were pretreated with (24.3%) than without (5.6%) Taxol. These results indicate that pretreatment of oocytes with Taxol before vitrification helps to reduce the damage induced by vitrification and is a potential way to improve the development of vitrified porcine oocytes.


2020 ◽  
pp. 161-169 ◽  
Author(s):  
Peter B. Best ◽  
Anabela Brandão ◽  
Doug S. Butterworth

Aerial counts of right whale cow-calf pairs on the south coast of South Africa between 1971 and 1998 indicate an annual instantaneouspopulation increase rate of 0.068 per year (SE = 0.004) over this period. Annual photographic surveys since 1979 have resulted in 901resightings of 550 individual cows. Observed calving intervals ranged from 2-15 years, with a principal mode at 3 years and secondarymodes at 6, 9 and 12 years, but these make no allowance for missed calvings. Using the model of Payne et al. (1990), a maximum calvinginterval of 5 years produces the best fit to the data giving a mean calving interval of 3.12 years (95% confidence interval: 3.07, 3.17). Thesame model produces an estimate for adult female survival rate of 0.983 (95% CI: 0.972, 0.994). The Payne et al. (1990) model is extendedto incorporate information on the observed ages of first reproduction of grey-blazed calves, which are known to be female. This allows theestimation of age at first parturition (median 7.88 years 95% CI 7.17, 9.29). Updates of estimates and confidence intervals for the otherdemographic parameters are: adult female survival rate 0.986 (0.976, 0.999); first year survival rate 0.913 (0.601, 0.994) and instantaneouspopulation increase rate 0.071 (0.059, 0.082). These biological parameter estimates are shown to be compatible with the observed increaserate of the population without the need to postulate immigration.


2020 ◽  
Vol 35 (10) ◽  
pp. 2262-2271
Author(s):  
D Cornet-Bartolomé ◽  
A Rodriguez ◽  
D García ◽  
M Barragán ◽  
R Vassena

Abstract STUDY QUESTION Is oocyte vitrification/warming as efficient and effective as using fresh oocytes in donation cycles? SUMMARY ANSWER IVF with vitrified donor oocytes is less efficient than using fresh oocytes, but its efficacy remains comparable to that of fresh cycles. WHAT IS KNOWN ALREADY Oocyte vitrification is used to preserve the reproductive potential of oocytes. A small number of randomized controlled trials carried out by experienced groups have shown that this technique provides fertilization, pregnancy, implantation and ongoing pregnancy rates comparable to those of fresh oocytes. However, large registry-based analyses have consistently reported lower live birth rates (LBRs) in cycles using vitrified oocytes. It is not clear whether this decrease may be due to the effect of vitrification per se on the oocytes or to the lower efficiency of the technique, as some of the oocytes do not survive after warming. STUDY DESIGN, SIZE, DURATION Retrospective cohort analysis of 1844 cycles of oocyte donation (37 520 oocytes), each donor in the study provided enough oocytes for at least one reception cycle with fresh oocytes (2561 cycles) and one reception cycle with vitrified oocytes (2471 cycles) from the same ovarian stimulation (sibling oocytes). Overall, 35 654 oocytes were considered in the analysis. All embryo transfers (n = 5032) were carried out between 2011 and 2017. PARTICIPANTS/MATERIALS, SETTING, METHODS Differences in reproductive outcomes after the first embryo transfer were evaluated using Pearson’s Chi-squared test and regression analysis adjusted for recipient’s age, BMI, sperm origin and state, day of embryo transfer, morphological score and number of transferred embryos. We performed two additional sub-analyses, to test whether the efficiency and/or effectiveness of vitrification/warming impacts reproductive results. One analysis included paired cycles where the same number of fresh and vitrified oocytes were available for ICSI (SAME sub-analysis), while the second analysis included those cycles with a 100% survival rate post-warming (SAME100 sub-analysis). MAIN RESULTS AND THE ROLE OF CHANCE Baseline and cycle characteristics of participants were comparable between groups. Overall, fertilization rates and embryo morphological scores were significantly lower (P &lt; 0.001) when using vitrified oocytes; moreover, vitrified oocytes also resulted in lower reproductive outcomes than sibling fresh oocytes using both unadjusted and adjusted analyses: ongoing pregnancy (32.1% versus 37.5%; P &lt; 0.001; OR 0.88, 95% CI 0.77, 1.00) and live birth (32.1% versus 31.9%; P = 0.92; OR 1.16, 95% CI 0.90, 1.49). However, when the efficiency of warming was taken into account, reproductive outcomes in recipients became comparable: ongoing pregnancy (33.5% versus 34.1%; P = 0.82; OR 1.11, 95% CI 0.87, 1.43) and LBR (32.1% versus 32%; P = 0.97; OR 1.15, 95% CI 0.89, 1.48). Moreover, after selecting only cycles that, in addition to having the same number of oocytes available for ICSI, also had 100% post-warming survival rate in the vitrified group, reproductive outcomes were also comparable between fresh and vitrified oocytes: ongoing pregnancy (34.8% versus 32.4%; P = 0.42; OR 1.32, 95% CI 0.98, 1.77) and live birth (32.9% versus 31.0%; P = 0.52; OR 1.27, 95% CI 0.95, 1.71), indicating that reproductive outcomes of these cycles are affected by the efficiency of the vitrification/warming technique performed rather than the oocyte damage due to the fast cooling process to which oocytes are subjected. LIMITATIONS, REASONS FOR CAUTION An open vitrification system was used for all cases, and oocyte vitrification/warming was performed by experienced embryologists with consistently high survival rates; caution must be exerted when extrapolating our results to data obtained using other open vitrification systems, closed vitrification systems or to IVF units with survival rates &lt;90%. WIDER IMPLICATIONS OF THE FINDINGS This is the largest cohort study comparing reproductive outcomes of vitrified and fresh sibling donor oocytes to date. We found that, when the number of oocytes available after warming is equal to the number of fresh oocytes, reproductive results including live birth are comparable. Consequently, the efficiency of vitrification must be taken into account to achieve the same reproductive outcomes as with fresh oocytes. We recommend implementing strict indicators of vitrification/warming efficiency in clinics and refining vitrification/warming protocols to maximize survival. STUDY FUNDING/COMPETING INTEREST(S) This work was supported by intramural funding of Clínica EUGIN and by the Secretary for Universities and Research of the Ministry of Economy and Knowledge of the Government of Catalonia (GENCAT 2015 DI 048). The authors declare no conflicts of interest. TRIAL REGISTRATION NUMBER N/A.


2008 ◽  
Vol 20 (1) ◽  
pp. 188 ◽  
Author(s):  
A. H. Sugulle ◽  
O. Dochi ◽  
H. Koyama

Prolonged exposure of oocytes to cryoprotectants causes cell injury, whereas a short exposure time results in insufficient permeation because of ice formation. This study was designed to determine the optimal pre-equilibration time and its effect on the survival rate of matured bovine oocytes after warming and on subsequent embryo development. Bovine COC were matured for 20 h in TCM-199 supplemented with 5% calf serum (CS) and 0.02 AU mL–1 of FSH at 38.5°C in 5% CO2 in air. Then, the COC were partially denuded. The oocytes were pre-equilibrated in 100 μL of vitrification solution 1 (VS1) containing 7.5% ethylene glycol (EG), 7.5% DMSO, and 20% CS in TCM-199 for 0, 1, 3, and 5 min. Then, the oocytes were moved through 100-μL drops of vitrification solution 2 (VS2) containing 30% EG, 30% DMSO, 0.5 m sucrose (Suc), and 20% CS in TCM-199 for 30 s, loaded into cryotops, and immersed into liquid nitrogen. Oocytes were warmed by plunging the cryotops into 1 m Suc in TCM-199 supplemented with 20% CS for 1 min, placed in 0.5 m Suc in TCM-199 supplemented with 20% CS for 3 min, and finally in TCM-199 supplemented with 20% CS alone for 5 min. Frozen–thawed semen from a single bull (5 × 106 spermatozoa mL–1) was used for fertilization. Zygotes were vortexed to remove the cumulus cells 18 h after fertilization and cultured in CR1aa for 9 days. Data were analyzed by the chi-square test. Results are presented in Table 1. There were no differences in the survival rates of the control and vitrified oocytes. The cleavage rate of controls at both 24 and 48 h was greater (P < 0.01) than that of vitrified oocytes. Among pre-equilibration times, the cleavage rate of 0, 1, and 3 min pre-equilibrations at both 24 and 48 h was greater than with the pre-equilibration time of 5 min (P < 0.01). With respect to blastocyst development, the control oocytes showed greater development rates than the vitrified oocytes (P < 0.01), whereas the development rates were lower with the pre-equilibration time of 5 min (P < 0.01) than with the other pre-equilibration times. In conclusion, the results indicated that matured bovine oocytes could survive after vitrification and subsequently develop into blastocysts after IVF. However, the pre-equilibration time before vitrification affects cleavage and blastocyst development; a longer exposure time resulted in lower blastocyst development. Table 1. Effect of pre-equilibration time on matured bovine oocytes survival and on subsequent embryo development


2005 ◽  
Vol 39 (3) ◽  
pp. 290-297 ◽  
Author(s):  
C A Trejo ◽  
M C Navarro ◽  
G D Ambriz ◽  
A Rosado

Preimplantation embryo development was studied in the golden hamster ( Mesocricetus auratus). Three groups of regularly cycling female hamsters were used: (I) 30 nulliparous young female (NYF) hamsters; (II) 24 nulliparous adult female (NAF) hamsters and (III) 30 multiparous adult female (MAF) hamsters. Female hamsters were mated with male hamsters of proven fertility. Only 15 min were allowed for mating. The moment of ejaculation was registered. Female hamsters were killed from 60 to 69 h after coitus. Corpora lutea were counted in both ovarian surfaces. Oviducts and uterine horns were flushed separately and embryo number, stage of development and distribution were recorded. Adult female hamsters, nulliparous and multiparous, had significant higher ovulation rates than NYF, but their reproductive efficiency was significantly lower. Preimplantation embryo development and transport were highly synchronous in NYF, but not in adults. Morulae were observed in NYF as early as 62–63 h after coitus. In adult female hamsters, significant numbers of morulae were found until 66–67 h. On the contrary, in NYF four-cell embryos were detected only until 60–61 h, while four-cell embryos were found until 64–65 h in NAF, and until 66–67 h in MAF. Embryo transport from the oviduct to the uterus is practically completed at 62–63 h after coitus in NYF, while it is evidently retarded in adult animals. In NYF all eight-cell embryos reached the uterus by 62 h after coitus. In adult female hamsters, both nulliparous and multiparous, a considerable number of eight-cell embryos fail to migrate into the uterus even at 67 h after coitus.


2013 ◽  
Vol 100 (3) ◽  
pp. S167
Author(s):  
S.H. Anderson ◽  
J. Gilchrist ◽  
J. Jones ◽  
E. Verrecchio ◽  
J.J. Orris ◽  
...  

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