• Title/Summary/Keyword: embryo culture

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Influence of the insemination method on the outcomes of elective blastocyst culture

  • Wang, Caizhu;Feng, Guixue;Zhang, Bo;Shu, Jinhui;Zhou, Hong;Gan, Xianyou;Lin, Ruoyun
    • Clinical and Experimental Reproductive Medicine
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    • v.44 no.2
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    • pp.85-89
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    • 2017
  • Objective: The aim of this study was to explore the effects of the insemination method on the outcomes of elective blastocyst culture. Methods: We retrospectively analyzed the outcomes of elective blastocyst culture performed between January 2011 and December 2014. Results: There were 2,003 cycles of conventional in vitro fertilization (IVF) and 336 cycles of intracytoplasmic sperm injection (ICSI), including 25,652 and 4,164 embryos that underwent sequential blastocyst culture, respectively. No significant differences were found in the female patients' age, basal follicle-stimulating hormone level, basal luteinizing hormone level, body mass index, number of oocytes, maturity rate, fertilization rate, or good-quality embryo rate. However, the blastocyst formation rate and embryo utilization rate were significantly higher in the conventional IVF group than in the ICSI group (54.70% vs. 50.94% and 51.09% vs. 47.65%, respectively, p<0.05). The implantation/pregnancy rate (IVF, 50.93%; ICSI, 55.10%), miscarriage rate (IVF, 12.57%; ICSI, 16.29%), and live birth rate (IVF, 42.12%; ICSI, 44.08%) were similar (p>0.05). No cycles were canceled due to the formation of no usable blastocysts. Conclusion: Although the fertilization method had no effect on clinical outcomes, the blastocyst formation rate and embryo utilization rate in the ICSI group were significantly lower than those observed in the conventional IVF group. Therefore, more care should be taken when choosing to perform blastocyst culture in ICSI patients.

Effect of Carbon and Nitrogen Source on Somatic Embryogenesis in Suspension Culture of Ligusticum chuanxiang Hort. (천궁의 현탁배양에서 탄소원과 질소원이 체세포배 형성에 미치는 영향)

  • Chae, Young-Am;Park, Sang-Un
    • Korean Journal of Medicinal Crop Science
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    • v.2 no.1
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    • pp.44-50
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    • 1994
  • This study was carried out to select the appropriate medium(especially, carbon and nitrogen source ) for somatic embryogenesis in order to develop the rapid mass production system in suspension culture of chuanxiang Hort. Suitable medium for somatic embryo formation was MS medium. The half strength MS medium was effective for somatic embryo development. Sucrose was the most effective carbon source for somatic embryo formation, however, production of somatic embryos was reduced at higher concentration of sucrose. Effects of suger was the same as sucrose. Somatic embryo formation was higher as the decrease of $NH_{4}NO_3$, and optimum ratio of $KNO_3\;:\;NH_4NO_3$ was 825 : 238mg /1. Regenerated plant was obtained in MS basal medium and survival late of plantlet was 60-70% after transplanted directly to the vermiculite.

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Addition of interleukin-6 to mouse embryo culture increases blastocyst cell number and influences the inner cell mass to trophectoderm ratio

  • Kelley, Rebecca L;Gardner, David K
    • Clinical and Experimental Reproductive Medicine
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    • v.44 no.3
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    • pp.119-125
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    • 2017
  • Objective: In vitro culture of preimplantation embryos is improved by grouping embryos together in a drop of media. Individually cultured embryos are deprived of paracrine factors; with this in mind, we investigated whether the addition of a single embryo-secreted factor, interleukin-6 (IL-6), could improve the development of individually cultured embryos. Methods: Mouse embryos were cultured individually in $2{\mu}L$ of G1/G2 media in 5% oxygen and supplemented with a range of doses of recombinant mouse or human IL-6. Results: Mouse IL-6 increased hatching at doses of 0.01 and 10 ng/mL compared to the control (93% and 93% vs. 78%, p< 0.05) and increased the total number of cells at a dose of 0.1 ng/mL compared to the control ($101.95{\pm}3.36$ vs. $91.31{\pm}3.33$, p< 0.05). In contrast, the highest dose of 100 ng/mL reduced the total number of cells ($79.86{\pm}3.29$, p< 0.05). Supplementation with human IL-6 had a different effect, with no change in hatching or total cell numbers, but an increase in the percentage of inner cell mass per embryo at doses of 0.1, 1, and 100 ng/mL compared to the control ($22.9%{\pm}1.1%$, $23.3%{\pm}1.1%$, and $23.1%{\pm}1.1%$ vs. $19.5%{\pm}1.0%$, p< 0.05). Conclusion: These data show that IL-6 improved mouse embryo development when cultured individually in complex media; however, an excess of IL-6 may be detrimental. Additionally, these data indicate that there is some cross-species benefit of human IL-6 for mouse embryos, but possibly through a different mechanism than for mouse IL-6.

Interactions between Recipients and Embyos affecting Pregnancy Rates in Bovine Embryo Transfer (수정란 이식에 있어서 수란우와 수정란의 상호작용이 수태율에 미치는 영향)

  • 이정호;박항균;조민희
    • Journal of Embryo Transfer
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    • v.1 no.1
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    • pp.76-80
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    • 1986
  • This study was carried out to investigate the inte actions between recipients and embryos to compare pregnancy rates in bovine embryo transfer, such as synchrony and stage of embryos, synchrony and quality of embryos, synchrony and side of uterine horn, and preservation time and stage of embryos. Fifty-two embryos were transferred by surgically to 42 Holstein heifers, 3 Holstein cows and 7 Korean native heifers from Feb., 1985 to June, 1986. The results were as follows: 1. In the trial of interactions between synchrony and embryo stage, recipients synchronired from- hours to + 12 hours in synchrony and embryos from morulac stage to advanced blastoctyst stages showed reason able pregnancy rate. 2. Excellant (A) and good (B) grade embryos showed good pregnancy rate, 81.8% and 73.3% respectively, but fair (C) grade embryos showed poor, 25% only at the same boundary of recipient synchrony. 3. More recipients had corpus luteum on the right ovary than the left, and also had better pregnancy rate (26/32 vs 13/20,81.3% vs 65%). 4. A good pregnancy rates (over 60%) were obtained with the embryo transferred 17hrs after flush at room temperature or field condition in culture medium.

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Chromosome Aberrations in Porcine Embryo Produced by Nuclear Transfer with Somatic Cell

  • Ah, Ko-Seung;Jin, Song-Sang;Tae, Do-Jeong;Chung, Kil-Saeng;Lee, Hoon-Taek
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.73-73
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    • 2002
  • Nuclear transfer (NT) techniques have advanced in the last years, and cloned animals have been produced by using somatic cells in several species including pig. However, it is difficult that the nuclear transfer porcine embryos development to blastocyst stage overcoming the cell block in vitro. Abnormal segregation of chromosomes in nuclear transferred embryos on genome activation stage bring about embryo degeneration, abnormal blastocyst, delayed and low embryo development. Thus, we are evaluated that the correlations of the frequency of embryo developmental rates and chromosome aberration in NT and In viかo fertilization (IVF) derived embryo. We are used for ear-skin-fibroblast cell in NT. If only karyotyping of embryonic cells are chromosomally abnormal, they may difficultly remain undetected. Then, we evaluate the chromosome aberrations, fluorescent in situ hybridization (FISH) with porcine chromosome 1 submetacentric specific DNA probe were excuted. In normal diploid cell nucleus, two hybridization signal was detected. In contrast, abnormal cell figured one or three over signals. The developmental rates of NT and IVF embryos were 55% vs 63%, 32% vs 33% and 13% vs 17% in 2 cell, 8 cell and blastocyst, respectively. When looking at the types of chromosome aberration, the detection of aneuploidy at Day 3 on the embryo culture. The percentage of chromosome aneuploidy of NT and IVF at 4-cell stage 40.0%, 31.3%, respectively. This result indicate that chromosomal abnormalities are associated with low developmental rate in porcine NT embryo. It is also suggest that abnormal porcine embryos produced by NT associated with lower implantation rate, increase abortion rate and production of abnormal fetuses.

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Comparison of static culture, micro-vibration culture, and micro-vibration culture with co-culture in poor ovarian responders

  • Hur, Yong Soo;Ryu, Eun Kyung;Yoon, San Hyun;Lim, Kyung Sil;Lee, Won Don;Lim, Jin Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.43 no.3
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    • pp.146-151
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    • 2016
  • Objective: This study was conducted to compare the effects of static culture, dynamic culture, and the combination of dynamic culture with specialized surfaces involving co-culture on human embryonic development. Embryos cultured using conventional static culture (SC) techniques served as a control group. We compared dynamic culture using micro-vibration culture (MVC) and micro-vibration with co-culture (MCoC), in which autologous cumulus cells were used as a specialized surface. Methods: We conducted a chart review of patients who were treated between January 2011 and November 2014 in order to compare embryonic development rates and pregnancy rates among the groups. Zygotes were cultured in micro-droplets, and embryos were subsequently selected for transfer. Some surplus embryos were cryopreserved, and the others were cultured for blastocyst development. A micro-vibrator was set at the frequency of 42 Hz for duration of 5 seconds per 60 minutes to facilitate embryo development. Results: No significant differences among the groups were present in patient's characteristics. However, the clinical pregnancy rates were significantly higher in the MVC group and the MCoC group than in the SC group. No significant differences were found in the blastocyst development rate between the SC group and the MVC group, but the blastocyst development rate in the MCoC group was significantly higher than in the SC and MVC groups. Conclusion: The clinical pregnancy rate was significantly increased by the application of micro-vibration to the embryonic cultures of poor responders. The blastocyst development rate was significantly increased by the application of MCoC to surplus embryos.

Effects of Glycerol Equilibration and Embryo Quality before Freezing and the Embryo Quality after Thawing in Mouse (생쥐에 있어서 Glycerol 평형단계 및 동결전 수정란 상태가 융해후 상태에 미치는 영향)

  • 조남기
    • Korean Journal of Animal Reproduction
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    • v.11 no.2
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    • pp.122-126
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    • 1987
  • This study was carried out to investigate the effects of the glycerol equilibration methods and embryo grades before freezing on the survival rate after thawing in mouse embryos. The results obtained from this study were as follows: 1. The number of embryos of grade I(Excellent), II(Good) and III(Fair) before freezing in this study was 97(27.4%), 160(45.2%) and 97(27.4%), respectively. 2. The average survival rate of frozen-thawed embryos in 3 and 5 steps glycerol equilibration was 66.7% and 64.1%, and the rate of transfererable embryos after culture was 68.9% and 69.0%, respectively. 3. Out of embryo grade I and II before freezing, the transferable rate after thawing was 75.2% and 48.1%, respectively, and grade I embryos before freezing was higher transferable rate than that of grade II.

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In vitro Production of Bovine Embryos - A Review

  • Rehman, N.U.;Sarwar, M.;Samad, H.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.9
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    • pp.1342-1351
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    • 2001
  • Over the years, the embryo transfer industry has grown from the simple collection & transfer of embryos into an advanced field of embryo biotechnology. Currently a large demand exists for bovine oocytes and early embryos in both research and commercial settings. Bovine embryos can now be produced in-vitro. Primary oocytes collected from antral follicles of abattoir - obtained ovaries can be induced to undergo the maturation process. In-vitor maturation system, however must ensure that the resulting oocyte is capable of undergoing normal fertilization and yields a zygote competent of developing to term after embryo transfer. Sperm preparation for IVF has improved with the use of heparine. The use of co-culture system has proved beneficial in circumventing the developmental block in IVM/IVF bovine embryos.

Embryo Rescue Efficiency Affected by Developmental Stages of Embryo and Medium Composition in Early-Ripening Peach (Prunus persica)

  • Sewon Oh;Byeonghyeon Yun;Se Hee Kim;Sang-Yun Cho;Namhee Jung;Kyung Ran Do;Kang Hee Cho;Hyun Ran Kim
    • Korean Journal of Plant Resources
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    • v.37 no.3
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    • pp.263-269
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    • 2024
  • Embryos of early-ripening peaches could not achieve physiological maturation or undergo abortion before harvest. Embryo rescue is an effective strategy to rescue embryos from early-ripening peaches. Thus, the current study was carried out to determine the appropriate developmental stage and optimal medium composition for embryo rescue in early-ripening peach. Development of open-pollinated 'Yumi' fruit was investigated from 20 to 90 days after full bloom (DAFB) to explore period occurring endocarp hardening. After endocarp hardening, embryo development was observed by light microscopes. Shoot and root meristems were observed at 65 DAFB and embryo size rapidly increased at 75 DAFB. Embryos collected at 75, 80, 85, and 90 DAFB were cultured on four media based on Driver and Kuniyuki (DKW) medium. Germination rate of embryos cultured on four media gradually increased from 75 to 90 DAFB and reached 100% at 90 DAFB. Notably, M3 medium (0.5 DKW supplemented with 6-benzylaminopurine (BAP) 1.0 ㎎/L) displayed the highest germination rate at 75 and 80 DAFB stages. Growth and development of shoot and root were pronounced in plantlet cultured at 90 DAFB stage. While delayed shoot growth was evident in plantlets cultured at 75, 80, and 85 DAFB stages, this retardation could be overcome through the application of growth regulators, particularly in M3 and M4 (0.5 DKW supplemented with BAP 1.0 ㎎/L and indole-3-butyric acid 0.5 ㎎/L) media. Remarkably, roots of plantlet grown in M4 medium exhibited limited elongation. In conclusion, germination rate of embryo and growth of embryo cultured plantlet can be enhanced by collecting seeds from early-ripening 'Yumi' at the 90 DAFB stage and conducting embryo culture using the M3 medium.