Analysis of related factors for blastocyst development
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Objective To explore related factors for blastocyst development during the in-vitro fertilization(IVF) and Intracytoplasmic sperm injection(ICSI) cycles.Methods D3 surplus embryos after embryo t ransfer or Vitrification from IVF/ ICSI cycles were cultured into blastocyst by the sequential method,then the blastocyst development were observed.Results A total of 7438 surplus embryos after embryo transfer or Vitrification were collected and formed 1382 high-quality blastulas(18.58%) af ter the sequential culture.At D3 cleavage stage,6-cell embryo had the significantly higher rate of high-quality blastula formation(10.34%) than 4 or 5-cell embryo(5.45%)(P=0.011);7-9cells embryo had the significantly higher rate of high-quality blastula formation(19.95%)than 6-cell embryo,≥10-cell embryo had the significantly higher rate of high-quality blastula formation(27.47%)than 7~ 9-cell embryo(P0.001).At D2 cleavage stage,4-cell embryo had the significantly higher rate of high-quality blastula formation(25.40%)than 3 or 5-cell embryo(10.26%,15.41%)(P0.001).At D2/ D3 cleavage stage,the embryo with I-IIgrade fragment had the significantly higher rate of high-quality blastula formation than the embryo with III-IV grade fragment(P0.001).Conclusion In-vitro blastocyst culture can have significant advantage to screen the high-quality embryo with development potential.The blastocyst formation has close relationship with D2/D3 embryo blastomere number and fragment.Keywords:
Blastula
Blastomere
Cleavage (geology)
Embryo quality
Vitrification
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Objective To evaluate the morphological assessment on embryonic developmental potentiality by investigating the blastula formation from day-3(D3) embryos. Methods A total of 1 185 infertile couples underwent in vitro fertilization- embryo transfer treatment were enrolled, of which the females were 21- 45 years old with mean age of 30.9 years old. Each blastocyst from naturally inseminated 8 148 D3 embryos was cultured in 25 μL droplet of culture medium, and highquality blastocyst formation rates were compared. Results The highest blastocyst rate was observed in 8-cell grade ⅠD3 embryos(76.49 %), which was significantly higher than that of 8-cell(35.10 %), 9-cell(68.12 %) or 9-cell(70.36 %) grade ⅠD3 higher rate of blastocyst formation in 8-cell grade ⅠD3 embryos was also observed in female 35-year-old as compared to those of ≥ 35-year-old(P 0.01). In subgroup ≥ 35-year-old, the blastocyst formation rate from D3 embryos was sequentially ordered as 8-cell grade Ⅰ(68.29 %) 8-cell grade Ⅰ(68.06 %) 7-cell grade Ⅰ(40.82 %) D3 embryos,with statistically significant differences among them(P 0.01). Conclusion It is demonstrated that 8-cell gradeⅠD3 embryo is optimal for cleavage-stage embryo transfer, however, 7- and 9-cell grade Ⅰ and Ⅱ embryos are the high-quality embryos.The criteria for D3 embryos in female ≥ 35-year-old needs to be further studied.
Blastula
Blastocyst Transfer
Cleavage (geology)
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The aim of this study was to identify the in vitro development stage at which the culture of a single or low number (n = 5 or 10) of oocytes/embryos could impair development in comparison with culture in group (n = 50). In the Experiment 1, it was confirmed that single in vitro embryo production yielded lower cleavage and blastocyst rates than in group (49.4 vs. 83.0%; 0% vs. 37.8%, respectively; p < 0.05). In Experiment 2 and 3, it was observed no effect on embryo development of culturing single or low number of oocytes during maturation and fertilization, respectively. In Experiment 4, it was observed a detrimental effect on blastocyst rate when cultured single or low number of embryos during post-fertilization in vitro culture (2.9; 10.2-10.8; 33.2% in single, low number of embryos (5-10), and controlgrouped, respectively; p < 0.05). In Experiment 5, it was observed that the last part of the culture period (day 3 onwards) seemed to be more affected by the low number of embryos placed in culture. In conclusion, post-fertilization culture, especially on days 3 to 7 after fertilization, seems to be the most important stage for embryo development on single and/or low number (5-10) of embryos culture.
In vitro maturation
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Objective: To compare the post-thaw survival rate and the success rate in continuing to grow to blastocyst of vitrified mouse embryos from different periods and different sources.Methods: The 2-cell,4-cell,8-cell period mouse embryos were obtained through fertilization in vitro and in vivo respectively.These embryos were vitrified and thawed.We got the rate of their survival and blastocyst formation.And,we took those fresh embryos as the control group.Results: The survival rate of blastocyst formation of post-thaw 2-cell,4-cell and 8-cell period embryos from fertilization in vitro and in vivo were 70.67%/65.33% and 72.06%/67.62%,88.00%/81.33% and 89.71%/83.82% and 93.33%/92.00% and 94.12%/94.12% respectively.The blastocyst formation rate in the phrase of 2-cell and 4-cell frozen embryos was lower than the control group(P0.05).No significant difference was found between the group of 8-cell frozen embryos and the control group.In addition,fertilization modes had no significant effect on the survival rate and the blastocyst formation rate(P0.05).Conclusion: Vitrification technology is an effective method in preserving mouse embryos.And the 8-cell period embryos might have the best cryopreservation effect.
Vitrification
Embryo cryopreservation
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Time-lapse imaging was used to establish the morphokinetics of equine embryo development to the blastocyst stage after invitro oocyte maturation (IVM), intracytoplasmic sperm injection (ICSI) and embryo culture, in oocytes held overnight at room temperature (22–27°C; standard conditions) before IVM. Embryos that developed to the blastocyst stage underwent precleavage cytoplasmic extrusion and cleavage to the 2-, 3- and 4-cell stages significantly earlier than did embryos that arrested in development. We then determined the rate of blastocyst formation after ICSI in oocytes held for 2 days at either 15°C or room temperature before IVM (15-2d and RT-2d treatment groups respectively). The blastocyst development rate was significantly higher in the 15-2d than in the RT-2d group (13% vs 0% respectively). The failure of blastocyst development in the RT-2d group precluded comparison of morphokinetics of blastocyst development between treatments. In any condition examined, development to the blastocyst stage was characterised by earlier cytoplasmic extrusion before cleavage, earlier cleavage to 2- and 4-cell stages and reduced duration at the 2-cell stage compared with non-competent embryos. In conclusion, this study presents morphokinetic parameters predictive of embryo development invitro to the blastocyst stage after ICSI in the horse. We conclude that time-lapse imaging allows increased precision for evaluating effects of different treatments on equine embryo development.
Reproductive immunology
Fertilisation
Theriogenology
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Sequential culture offers the early embryo with the nutrition in the different developmental stages.In the experiments,the rabbit pronuclear embryo was cultured with 10% FBS+RPMI1640 and 10% FBS+mRPMI1640 by sequential culture,and the control is with 10% FBS+RD by single culture.RESULTS:At 72 h of culture,the ratios of 8-cell,morula and blastocyst are not significant different(P0.05) in the both culture,but 10.3% fertilized oocytes has been degenerated in the single culture with RD,which is significant difference to the sequential culture(10.3% VS 0,P0.05).At 168 h of culture,the ratio of attachment,outgrowth,morula,hatched embryos is 59.7% VS 41.4%,43.1% VS 22.4%,100% VS 89.7%,33.3% VS 5.2%,respectively in both way.The two formers are significant different(P0.05),the two latters are significant different very much(P0.01) in both culture way..The number of the blastocyst cell is 124.6±6.36 in sequential culture and 118.2±5.25 in RD,which is significant different(P0.05).The results showed that sequential culture can overcome the developmental block in rabbit early embryo,support the embryo to growth and development,improve the quality of embryo,and promote embryo to hatching and implantation.
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Blastomere
Cleavage (geology)
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The absence of reliable markers for the identification of viable embryos for transfer at the early cleavage stage is likely to contribute to the generally low implantation rates and high incidence of multiple gestation in IVF treatment. In this study, we investigate the relationship between timing of first cleavage and the incidence of blastocyst formation in vitro.Couples (n = 70) with at least one embryo remaining after transfer were included in the analyses. All embryos (n = 579) were examined for early cleavage at 25 h after insemination. Following embryo transfer, the remaining embryos (n = 426) were cultured until day 7 of development, and assessed for blastocyst formation.Eighty-five embryos (14.7%) cleaved to the 2-cell stage within 25 h of insemination; 26 of these were selected for transfer on day 2. Of the 59 embryos remaining in culture, 19 (32.2%) developed to the blastocyst stage; this was a significantly higher number than was observed in embryos (61/367; 16.6%) that failed to cleave within 25 h of insemination (P < 0.01). Within these two groups of embryos the proportion of hatched blastocysts was 11/59 (18.6%) and 26/367 (7.1%) respectively (P < 0.005).These findings indicate that early cleavage is indicative of increased developmental potential in human embryos and may be useful as an additional criterion in the selection of embryos for transfer.
Cleavage (geology)
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Vitrification
Inner cell mass
Embryo cryopreservation
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Objective To investigate the feasibility of screening viable embryos from poor quality embryos by extending blastocyst culture. Methods 538 D3 poor quality embryos from 142 cases IVF /ICSI treatment were collected after embryo transfer or freezing,and observe the blastocyst formation by blastocyst cultured for up to the sixth days( D6). The embryos were divided into four groups( 4 cells grade Ⅰ /Ⅱ /Ⅲembryos,5 cells grade Ⅰ /Ⅱ /Ⅲ embryos,6 ~ 9 cells grade Ⅲ embryos and exceed 9 cells grade Ⅰ /Ⅱ /Ⅲembryos) according to different blastomere and grade,and then compared the blastula development. Results The blastula formation rate and the high-quality blastula rate from D3 poor quality embryos were 65. 6% and 51.3%,the blastula formation rate of the four groups were 55. 6%,56. 6%,70. 4% and 66. 7%,there were significant difference among the four groups( P 0. 05). And the high-quality blastula rate were 8. 9%,32. 6%,61. 0% and 63. 3%,there were significant difference among the four groups( P 0. 05). Conclusions Extending blastocysts cultured can select the developmental potential embryos from those poor quality embryos remnant effectively,and make better use of the embryos.
Blastula
Blastomere
Embryo quality
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Objective: To study the clinical effects of blastocyst culture technique on the outcome of in vitro fertilization-embryo transfer(IVF-ET). Methods: 292 ova from 24 patients were fertilized and sequentially cultured in Quin’s media plus 10% human serum albumin(HSA)in vitro. In 24 IVF-ET cycles, transplantation was performed at the blastocyst stage. Conventional culture that the transplantation was done on the third day after fertilization was carried out in 28 IVF-ET cycles. The fertilization rate, cleavage rate, the numbers of embryo transfered (NET) and the pregnancy rate were compared between these two groups. Results: The NET were significantly smaller in blastocyst culture group than those in conventional culture group(P0.001). Conclusion: Blastocyst culture is more suitable for selecting good embryos to transfer, which can increase the pregnancy rate and decrease the multiple pregnancy rate.
Blastocyst Transfer
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