• Title/Summary/Keyword: Embryo morphology

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Comparison of In Vitro Embryo Production with Follicular Oocytes Collected by Aspiration and Slicing in Korean Native Cows (한우 난포란의 채란방법에 따른 체외수정란의 생산효율)

  • 이경미;곽대오;송상현;최양석;김윤연;강다원;하란조;윤창현;박충생
    • Journal of Embryo Transfer
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    • v.11 no.3
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    • pp.249-258
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    • 1996
  • To improve the efficiency of in vitro production of embryos with follicular oocytes in Korean Native cows, the recovery rates, in vitro maturation, fertilization and development, and the time required for collecting and processing oocytes by aspiration with or without slicing were evaluated comparatively. The ovaries were obtained from a local abattoir and placed in physiological saline at 25~28$^{\circ}C$ and brought to the laboratory within 3 hrs. The oocytes were collected by aspiration of follicles(2~6mm) with or without slicing ovaries after aspiration, and classified into Grade I, Grade II, Denuded, Expanded oocytes by the morphology of cumulus cells attached and the homogeneity of cytoplasmic granules. Also the time required for each step of collecting and processing oocytes were measured. The cumulus cells were removed in some Grade I oocytes to measure their size and nuclear configuration before and after in vitro maturation. The Grade I oocytes were matured in vitro(IVM) for 24 hrs. in TGM-199 supplemented with 35$\mu$g /ml FSH, 10$\mu$g /ml LH, 1 $\mu$g /ml at 39$^{\circ}C$ under 5% C02 in air. They were fertilized in vitro(IVF) by epididymal spermatozoa treated with heparin for 24hrs. and then the zygotes were cocultured in vitro (IVC) with bovine oviductal epithelial cells for 10 days. The results obtained were as follows: The number of oocytes recovered per ovary was averaged 6.6 by aspiration and 11.2 by slicing post aspiration, which summed to 17.8. The number of Grade I oocytes recovered per ovary was averaged 3.1 by aspiration and 3.6 by slicing, which summed to 6.7. The percentage of Grade I to total oocytes recovered was significantly(P<0.05) higher as 48.0 % in aspiration than 31.6% in slicing post aspiration. The time requlred for recovering a Grade I oocyte by aspiration and slicing was 1.1 and 2.5 min, respectively. The mean diameter of Grade I oocytes by aspiration and slicing was similar as 148.7 and 151.5$\mu$m, respectively. The percentage of Metaphase II stage oocytes after IVM for 24 hours was significantly (P

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The impact of post-warming culture duration on clinical outcomes of vitrified-warmed single blastocyst transfer cycles

  • Hwang, Ji Young;Park, Jae Kyun;Kim, Tae Hyung;Eum, Jin Hee;Song, Haengseok;Kim, Jin Young;Park, Han Moie;Park, Chan Woo;Lee, Woo Sik;Lyu, Sang Woo
    • Clinical and Experimental Reproductive Medicine
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    • v.47 no.4
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    • pp.312-318
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    • 2020
  • Objective: The objective of the study was to compare the effects of long-term and short-term embryo culture to assess whether there is a correlation between culture duration and clinical outcomes. Methods: Embryos were divided into two study groups depending on whether their post-warming culture period was long-term (20-24 hours) or short-term (2-4 hours). Embryo morphology was analyzed with a time-lapse monitoring device to estimate the appropriate timing and parameters for evaluating embryos with high implantation potency in both groups. Propensity score matching was performed to adjust the confounding factors across groups. The grades of embryos and blastocoels, morphokinetic parameters, implantation rate, and ongoing pregnancy rate were compared. Results: No significant differences were observed in the implantation rate or ongoing pregnancy rate between the two groups (long-term culture group vs. short-term culture group: 56.3% vs. 67.9%, p=0.182; 47.3% vs. 53.6%, p=0.513). After warming, there were more expanded and hatching/hatched blastocysts in the long-term culture group than in the short-term culture group, but there was no significant between-group difference in embryo grade. Regarding pregnancy outcomes, the time to complete blastocyst re-expansion after warming is shorter in women who became pregnant than in those who did not in both culture groups (long-term: 2.19±0.63 vs. 4.11±0.81 hours, p=0.003; short-term: 1.17±0.29 vs. 1.94±0.76 hours, p=0.018, respectively). Conclusion: The outcomes of short-term culture and long-term culture were not significantly different in vitrified-warmed blastocyst transfer. Regardless of the post-warming culture time, the degree of blastocyst re-expansion 3-4 hours after warming is an important marker for embryo selection.

Apoptosis in CHSE-214 Chinook Salmon Embryo Cells Infected with Hirame Rhabdovirus (HIRRV)

  • Sung Han-Gi;Kim Yeong-Jin;Jung Sung-Ju;Choi Won-Chul;Jung Tae-Sung;Choi Tae-Jin;Oh Myung-Joo
    • Fisheries and Aquatic Sciences
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    • v.5 no.1
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    • pp.32-35
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    • 2002
  • In this study, we investigated the mechanism of cell death in rhabdovirus-infected cells, chinook salmon embryonic cell line (CHSE-2l4) infected with hirame rhabdovirus (HIRRV). Studies using light microscopy, fluorescence microscopy, TUNEL method, electron microscopy, and agarose gel electrophoresis revealed changes in the cell morphology and DNA fragmentation indicative of apoptosis in early infection. It was observed that HIRRV induced apoptosis as well as necrosis in infected cells.

Interspecific Hybrid of Nicotiana trigonophyllo X N. tabacum through in vitro Culture of Fertilized Ovules (배주배양에 의한 Nicotiana trigonophylla와 N. tabacum의 종간잡종 육성)

  • 최상주;이승철
    • Journal of the Korean Society of Tobacco Science
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    • v.13 no.2
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    • pp.52-58
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    • 1991
  • Interspecific cross between Nicotiana trigonophylla and N. tabacum cv. BY4 is highly sterile because of abnormal ovule and embryo development. In vitro culture of excised N. trigonophylla ovules after polination by N. tabacum allows significant numbers of hybrid embryos to develop into mature plants. Total yield of seedlings and number of normal seedlings were produced following in vitro culture of individual fertilized ovules of N. trigonophylla X N. tabacum at four days post-pollination on B5 medium containing 6% sucrose. Hybrids were uniform in morphology and peroxidase isozyme composition and the majority were cytologically stable: flower characteristics were generally intermediate between those of the parents. All hybrids evaluated were self-sterile.

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Rearrangement of Korean Terms for Insect Head Morphology (곤충 머리 부위에 대한 우리말 용어 재조명)

  • Lee, Young In
    • Korean journal of applied entomology
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    • v.57 no.4
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    • pp.279-285
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    • 2018
  • Different English terms indicating a same part in insect head were arranged according to position and function, and their corresponding different Korean terms were suggested. The terms include lines on head surface, head regions of embryo, external and internal skeletons, internal parts of mouth, long mouths, external parts and basic segments of antenna, antennal shapes, and hairs on surface.

Preservation and Transfer of Bovine Embryos by Vitrification Method

  • Lee, S.Y.;J.S. Yu;D.S. Chung;Park, C.K.
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.134-134
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    • 2003
  • Bovine embryos produced by in vitro maturation, feretilization and development was examined for presevation and transfer. The fertilization medium used BO medium with 5 mM/$m\ell$ caffeine and 10$\mu\textrm{g}$/$m\ell$ heparin and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to 1$\times$$10^{6}$ cells/$m\ell$ motile sperm during fertilization in vitro. At 8~10 hrs after insemination, the oocytes were transferred into CR1aa medium and cultured for 7 days. Embryos were preserved by vitrification method for transfer. When the embryos of early, blastocyst and expanded blastocyst stages were frozen-thawed, the proportions of embryos with normal morphology 83.6, 88.1 and 85.2%. (중략)

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Male Reproductive Toxicity of DA-125, a New Anthracycline Anticancer Agent, in Rats (수컷랫드에 있어서 새로운 안트라사이클린계 항암제 DA-125의 생식독성 연구)

  • 김종춘;김갑호;신호철;정문구
    • Toxicological Research
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    • v.14 no.2
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    • pp.193-203
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    • 1998
  • The toxicity of DA-125. a new anthracycline anticancer agent, on the male reproductive system was studied in Sprague-Dawley rats. Forty male rats were rando$m\ell$y assigned to Jour groups with ten rats in each group and given single intraveneous doses of DA-125 at dose levels of 0. 12.5. 25. and 50 mg/kg body weight. On day 56 after treatment the animals were allowed to mate. and their male reproductive Junctions and organs were examined in detail. Copulated females were sacrificed on day 20 of gestation for examination of embryo-fetal development. One out of ten rats in the 50 mg/kg group died on day 12 after treatment. Clinical signs such as emaciation. sedation, anorexia. swelling. dark material around eye. alopecia. and diarrhea were observed in the 25 and/or 50 mg/kg groups. Reduction in the body weight gain. decrease in the absolute weights of testes. epididymis and seminal vesicles. and/or decrease in the number of testicular sperm heads were also found. Although histopathological changes such as atrophy of seminiferous tubules. loss or decrease of spermatogenic cells. exfoliation of spermatogenic cells. vacuolization of Sertoli cells. decrease of sperm. and/or increase of necrotic spermatogenic cells in epididymal ducts were observed. no adverse effects on the motility and morphology of epididymal sperm. copulation index. fertility index. and embryo-fetal development were detected in the 25 and 50 mg/kg groups. There were no evidences of male reproductive toxicity in the 12.5 mg/kg group. These results show that single intravenouse doses of DA-125 produce significant dose-related testicular atrophy. histopathological changes. and oligozoospermia in rats and $LD_{10}$ for DA-125 appears to be 50 mg/kg body weight.

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In Vitro Development of Preantral Follicles Isolated from Juvenile Mice (약령 마우스에서 분리한 난포난자의 체외발생)

  • 이현주;김선영;김기동;이상호;송해범
    • Journal of Embryo Transfer
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    • v.17 no.3
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    • pp.203-209
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    • 2002
  • The aim of this study was to assess the developmental capacity of oocytes maturated in vitro after 10 days of culture when the preantral follicles were isolated from juvenile mice 10- and 20-day old, respectively, and to develop in vitro culture system that observed a view to morphology of follicles and nucleus maturation of oocytes. The antral-like cavities became formation after 6 days of culture in follicle isolated from 10- and 20-day old mice. The number of follicles were 21.5 and 33.3 in ovary isolated from 10- and 20-day old mice, respectively. The diameters of oocytes were 51.85 and 57.50 ${\mu}{\textrm}{m}$ before culture and were grew 55.95 and 63.11 ${\mu}{\textrm}{m}$ after culture for 10 days, in follicles isolated from 10- and 20-day old mice, respectively. The observation rates up to the M II and from GV to M II were 4.3 and 22.1%, and 14.5 and 61.1% after culture for 10 days in follicles isolated from 10- and 20-day old mice, respectively.

A Simple Embryonic Stem Cell-Based in vitro Differentiation System That Recapitulates Early Erythropoietic Events in the Mouse Embryo (생쥐 배아에서의 초기 적혈구 분화를 재현 할 수 있는 배아주 세포에 기초한 간단한 시험관내 분화체계)

  • 김철근
    • The Korean Journal of Zoology
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    • v.39 no.3
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    • pp.239-247
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    • 1996
  • An embryonic stem (ES) cell-based in vitro model system was examined to determine whether a simple differentiation of embryoid bodies (EB) in the suspension medium is useful to dissect early erythropoiesis. Characteristics of the differentiating EBs were monitored for their differentiation potential to generate hematopoietic cell types by general morphology, benzidine staining and two-step colony assays, and expressivity of several erythroid marker genes by the RT-PCR analysis for total cellular RNA prepared from the differentiating EBs. Every ematopoietic lineage cells were generated from the differentiating EBs with reproducible frequencies, similar to the other sophisticated differentiation protocols. Furthermore, the globin gene switching in differentiating ES cells paralleled the sequence of events found in the mouse embryo, and such that their expression was activated by at least 12 hrs later than those of erythroid-specific transcription factors, GATA-1 and Tal-1 The erythropoietic differentiation program initiated reproducibly and efficiently in this simple differentiation system in a suspension culture, such that this system may be useful for dissection of the molecular events of early erythropoiesis.

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Duration of Preservation Affect the Quality of Chilled Black Bengal Buck Semen

  • Pradhan, Md. Gulshan Anowar;Rahman, Md. Saidur;Kwon, Woo-Sung;Mishra, Dipendra;Kamal, Md. Mostofa;Bhuiyan, Mohammad Musharraf Uddin;Shamsuddin, Mohammed
    • Journal of Embryo Transfer
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    • v.28 no.2
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    • pp.113-119
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    • 2013
  • The study focuses on the quality assessment of Black Bengal buck semen preserved at chilled condition. In this in vitro trial, collected semen from Black Bengal bucks was preserved at chilling temperature ($4{\sim}5^{\circ}C$) in tris-glucosecitrate yolk medium of 1:5 ratios for four days. Artificial Vagina (AV) method was utilized to collect semen from buck. General evaluation of semen includes the color, mass activity and density were measured by direct visual examination. However, computer-assisted sperm analysis (CASA) and phase contrast microscopy were used to figure out the motility (%), hyper-activated (HYP) motility (%) and number of abnormal spermatozoa (%) initially, and at every 24 h intervals. The result revealed that spermatozoa preserved at chilling temperature showed significantly (P<0.05) lower motility and HYP motility with the progression of preservation. The number of phenotypically abnormal spermatozoa significantly (P<0.05) increased following preservation. Although significant positive correlation (r=0.945; P<0.05) was existed between % motile and % HYP motile spermatozoa however, the % of morphologically abnormal spermatozoa was negatively correlated with % motile (r=-0.997; P<0.05) and % HYP motile spermatozoa (r=-0.946; P<0.01). Therefore, we concluded that the quality of chilled semen progressively losses its viability and doesn't remain useable after certain period of preservation with respect to its motility and morphology.