• Title/Summary/Keyword: Donor Development

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Developmental Potential of Interspecies Nuclear Transferred Embryos using Mouse Embryonic Fibroblast In Vitro

  • B.S.Koo;Yoon, J.I.;Son, H.Y.;Kim, M.G.;Park, C.H.;Lee, S.G.;Lee, Y.I.;Lee, C.K.
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.109-109
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    • 2003
  • Even though success in birth of live offspring from nuclear transfer(NT) using somatic cells in many species, detailed information on processes or mechanisms of development are not well known. Cytoplasm of bovine oocyte has been known to support the development of nuclear transferred embryos using nuclear donor cells from different species. Therefore, interspecies NT might be used to find answers of some questions in basic aspect of nuclear transfer In this study, we examined the developmental potential of reconstructed embryos when bovine oocyte as a cytoplasm recipient and mouse embryonic fibroblast as a nuclear donor were used. The nuclear transfer units were aliocated in Group 1 (murine block media and normal media) and Group 2. (bovine block media and normal media). NT units were not blocked at 2-cell stage regardless of types of medium. On mouse media, poor development of interspecies NT units was observed compared to bovine media. However, as NT units cultured in bovine normal medium, embryos developed over 8-cell stage. Further studies performed to increase the developmental rate in condition of antioxidant treatment. Despite low development, bovine-murine interspecies nuclear transferred embryos could develop to blastocysts and they showed that blastocyts rate of antioxidant group was superior to those of non-antioxidant group. Next, we investigated gene expression pattern which is carried out for zygotic activation. The Xist gene is expressed in female mouse embryo after zygotic activation of 4-cell stage. But interspecies nuclear transferred embryos do not express Xist gene at 4-cell stage. As a result, it is suggested that the bovine cytoplasm controls the early preimplantation development in interspecies NT However, the development of later stages might require genomic control from transferred donor nucleus. Therefore, even though the involvement of several other factors such as mitochondrial incompatibility, effective development of embryos produced by interspecies NT requires proper genomic activation of donor nucleus after overcoming the cytoplasmic control of recipient oocytes.

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Development of Somatic Cell Nuclear Transfer Embryos Following Donor Cell Type and Cell Treatment in Cattle (Donor 세포의 종류 및 세포처리에 따른 소 체세포 핵이식란의 체외발육)

  • 손준규;박정준;박춘근;양부근;김정익;정희태
    • Reproductive and Developmental Biology
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    • v.28 no.1
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    • pp.1-6
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    • 2004
  • This study was conducted to investigate the effects of donor cell type, individual, passage number and trypsinization time on the in vitro development of bovine somatic cell nuclear transfer embryos. Three cell types (skin, muscle and cumulus cells) and cells from 3 individuals were used for nuclear transfer. Cell were passaged by 5, 15 or 30 times, and cell were trypsinized for 1 or 3 min before injection. Nuclear transfer were performed by conventional fusion method. Development rates to the blastocyst stage were not significantly different among three cell types (16.5∼23.9%) and individuals (16.4∼19.5%). Blastocyst formation rate of cloned embryos reconstituted with cells at passage 30 (5.8%) was significantly lower than those of embryos reconstituted with 5- and 15-passaged cells (25.3 and 23.5%, respectively, P<0.05). The rate of embryos developed to the blastocyst stage was higher in embryos reconstituted with cells trypsinized for 1 min (30.7%) compared to embryos reconstituted with cells trypsinized for 3 min (P<0.05). The result of the present study indicates that different donor cell types and individuals used in this study did not affect the development of cloned bovine embryos. However, passage number and trypsinization time of donor cells affect the in vitro development of cloned bovine embryos.

Effect of Nuclear Transfer Methods on In Vitro Development of Reconstituted Bovine Embryos II. Effect of Electric Voltage and In Vitro Produced Donor Embryo Quality on Fusion and In Vitro Development (소에서 핵이식 방법이 재구축배의 체외발달에 미치는 영향 II. 통전전압과 체내생산 공핵배의 질에 따른 융합과 체외발달)

  • 정영채;김창근;송학웅;정영호;윤종택;이종완;김흥률;김광식
    • Korean Journal of Animal Reproduction
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    • v.20 no.4
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    • pp.467-472
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    • 1997
  • This study was conducted to determine the optimal DC voltage for NT of in vivo donor embryo nuclei and investigate the effect of donor embryo guality on fusion and in vitro development of NT embryos . Recpient oocytes were enucleated 25~27h after IVM and further cuitured for 18~20h prior to fusion for oocyte aging. Donor embryos of molura stage were recovered from superovulated heifers and classified l into good and low quality group. Their nuclei were transferred in to the emucleated oocytes 42~44h post-IVM and fused 43-45h post-IVM with a single 0.75kV /cm or 1.0kV /cm DC voltage for 70${\mu}\textrm{A}$sec. The fusion rate of oocytes was not different between two DC voltages. However, the cleavage and M + B developmnent was more high at 1.0kV /cm DC voltage and the proportion of M+B was 19.0% at 0.75kV /cm DC and 29.4% at 1.0kV /cm DC voltage. Donor embryo qualtiy did not greatly affect the fusion and cleavage of NT oocytes, but none of NT embryos derived from low embryo quatity reached the morula stage. The results indicate that the most suitable DC v voltage for electrofusion of in vivo donor muclei was a single 1.0kV /cm DC voltage and donor embryo quality was an important factor affecting the development in vitro of NT embryos.

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Nucleo-cytoplasmic Interactions of Bovine Oocytes and Embryos Following Nuclear Transplantation (핵이식에 의한 소 난자 및 초기배의 핵-세포질의 상호작용에 관한 연구)

  • 김정익;양부근;정희태
    • Korean Journal of Animal Reproduction
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    • v.17 no.4
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    • pp.287-294
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    • 1994
  • This study was to investigate the effects of electrofusion, activation and developmental stage of donor embryos on in vitro development of nuclear transplant bovine embryos. A single blastomere nucleus from 8-cell to morula stage embryos produced by in vitro fertilization(IVF) was transferred into a recipient oocyte enucleated at 23∼25 h after in vitro maturation(IVM) or into a recipient oocyte enucleated and cultured for 14∼15 h. In one experiment the nuclear transplant embryos were subjected to additional activation treatments. Fusion rate of nuclear transplant eggs was high at direct current(D.C) voltages of 1.0 and 1.5 kV/cm 991.5 and 93.3%, respectively), but decreased at 2.0kV/cm (81.8%). Additional activation treatments by electric pulases or 7% ethanol did not affect the cleavage and development of nuclear transplant embryos. Development of nuclear transplant embryos slightly increased by delayed nuclear transfer and fusion (42∼43 h after IVM). With this system, blastocysts were obtained from transfer of 8-cell to morula stage donor nuclei (9.6%∼2.4%). The result of this study suggests that nucleo-cytoplasmic interactins, expecially activation of ooplast are very important for the development of nuclear transplant embryos, and donor cell stage does not affect the development of nuclear transplant embryos.

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The Effects of Donor Cell Type and Culture Medium on in vitro Development of Domestic Cat Embryos Reconstructed by Nuclear Transplantation

  • Fahrudin, Mokhamad;Otoi, Takeshige;Karja, Ni Wayan Kurniani;Murakami, Masako;Suzuki, Tatsuyuki
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.8
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    • pp.1057-1061
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    • 2001
  • In this study we explored the possibility of performing nuclear transfer in the domestic cat and assessed the ability of different culture media to support in vitro development of reconstructed cat embryos. Donor somatic cells were derived from cultured cumulus cells or explants of oviduct tissue, and recipient cytoplasts from in vitro matured oocytes. A higher percentage of cleavage (84.6% and 86.5%) and development to the morula stage (35.9% and 44.2%) was found when reconstructed embryos receiving cumulus or oviduct cells were cultured in MK1 medium, compared with those cultured in CR1aa (58.7% and 72.5%, 13.8% and 13.6%, respectively). There was no significant difference between MK1 and CR1aa media with respect to the proportion developing to the blastocyst stage (15.4% and 17.3% vs 6.8% and 8.6%, respectively, p>0.05). There was no significant effect (p>0.05) of donor cell type (cumulus and oviduct cells) on the rates of fusion (65.0% and 52.5%), cleavage (84.6% and 86.5%), development to the morula (35.9% and 44.2%), and blastocyst (15.4% and 17.3%) stages when reconstructed embryos were cultured in MK1 medium. Similar results were found for the reconstructed embryos cultured in CR1aa medium. These results show that culture medium has a significant impact on the early development of reconstructed cat embryos, whereas donor cell type does not have a significant effect.

Basic and Prospective Aspects on Allelopathic Research (알레로파시 연구의 기초와 전망)

  • 길봉섭
    • Proceedings of the Korean Society of Environment and Ecology Conference
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    • 2005.04a
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    • pp.3-14
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    • 2005
  • To investigate phytotoxic substances in some donor plants and their biological activities, seed germination and seedling growth of receptor plants were examined at different concentrations of aqueous extracts and essential oils of the donor plants. Germination of some receptor species was inhibited by the extracts, while seedling growth was decreased to a lesser degree than in the germination test. Germination, seedling growth and dry weight growth of Achyranthes japonica grown in pot were proportionally inhibited by the extracts. Volatile substances emitted some donor plants caused inhibitory effects in the germination and seedling growth of the receptor species. Essential oil of the plants extracted by Karlsruker's apparatus Inhibited growth of microorganisms, callus growth and root hair development of receptor plants. The cortical cells at the root tips of Latuca sativa treated with essential oils showed contraction of the cytoplasm, resulting in plasma membranes becoming detached from the cell walls and the cells metamorphosing irregularity. Accumulation of lipid granules Inside contracted cytoplasm and degeneration of mitochondrial cristae were also observed. The GC/MS method was employed for analysis and identification of allelochemicals from donor plants. Sixty-one chemical substances such as camphene, cineole etc. were identified from essential oils of Artemisia argyi.

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STUDIES ON PRODUCTION AND EFFICIENT UTILIZATION OF LIVESTOCK EMBRYOS BY IN VITRO FERTILIZATION AND MICROMANIPULATION IV. NUCLEAR TRANSPLANTATION AND ELECTROFUSION FOR CLONING IN BOVINE FOLLICULAR OOCYTES

  • Chung, Y.C.;Kim, C.K.;Song, X.X.;Yoon, J.T.;Choi, S.H.;Chung, Y.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.8 no.6
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    • pp.641-645
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    • 1995
  • This study was conducted to develop a method for production of nuclear transplant bovine embryos using in vitro-matured (IVM) oocytes and to examine the effect of different conditions of electrofusion on fusion rate and developmental capacity of donor nucleus transplanted to enucleated oocytes. Eight- to sixteen-cell embryos derived from oocytes matured and fertilized in vitro used as donor blastomeres and IVM oocytes were used as recipient oocytes. Oocytes were enucleated immediately after 23-24 h IVM and then reconstituted with a donor blastomere in two different micromanipulation media. Fusion rate and subsequent development of the reconstituted oocytes was compared under the different electric stimuli and recipient oocyte ages. Success rate of enucleation was significantly higher in TCM-199 medium containing FCS than in DPBS. The high fusion rate(75-94%) and development (6.4-14.8%) to morulae and blastocyst (M + B) were obtained from 0.6-0.75 kV/cm DC voltage, although total cleavage was not different among the electric pulses. Most optimal condition of electric stimulation for fusion and development was 1 DC voltage of 0.75 kV/cm, in which 80.5% of oocytes were fused, 80.0% and 31.7% of which was cleaved and developed to M + B, respectively. No M + B was obtained from 1.2 kV/cm DC voltage regardless of pulse frequency. Recipint oocyte age at electrofusion greatly affected the cleavage and subsequent development to M + B, showing high rate at 40-41 h oocyte maturation. These results suggest that a suitable condition of electrofusion for donor nuclei derived from IVF may be 1-2 DC pulses of 0.7 kV/cm for $70{\mu}sec$ and that processing of a transplanted nucleus in IVM oocytes may be affected by maturation age of recipient oocytes.

The Study on Phase Transition Pressure of Donor doped Pb(Zr0.52Ti0.48)O3 Ceramics with Diamond Anvil Cell (다이아몬드 엔빌 셀을 이용한 Donor doped Pb(Zr0.52Ti0.48)O3 세라믹스의 상전이 압력 연구)

  • Cho, Kyung-Ho;Ko, Young-Ho;Seo, Chang-Eui;Kim, Kwang-Joo
    • Journal of the Korean Ceramic Society
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    • v.48 no.5
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    • pp.471-478
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    • 2011
  • Investigations of crystal structure and phase transition of $Pb(Zr_{0.52}Ti_{0.48})O_3$ ceramics doped with A-site substitution impurity (La, Nd) or B-site substitution impurity (Sb, Nb) at 2 mol% concentration were carried out. X-ray diffraction patterns of impurities doped $Pb(Zr_{0.52}Ti_{0.48})O_3$ ceramics have been measured at pressures up to ~5 GPa with diamond anvil cell and synchrotron radiation. The patterns were obtained at room temperature using methanol-ethanol mixture as pressure-transmitting media. In order to refine the crystal structure, Rietveld analysis has been performed. The structures of impurities doped $Pb(Zr_{0.52}Ti_{0.48})O_3$ ceramics are tetragonal in space group P4mm at ambient pressure and are transformed into a cubic phase in space group Pm$\bar{3}$m as the pressure increases. In this study, when A-site substitution donor $La^{3+}$ or $Nd^{3+}$ ion was added to $Pb(Zr_{0.52}Ti_{0.48})O_3$ ceramics, the phase transition phenomena showed up at the pressure of 2.5~4.6 GPa, but when B-site substitution donor $Nb^{5+}$ or $Sb^{5+}$ ion was added to it, the phase transition appeared at relatively lower pressure of 1.7~2.6 GPa.

Recipient Countries' Financial Development and the Effectiveness of ODA (금융시장발전과 공적개발원조의 효과성: 양자간·다자간 원조를 중심으로)

  • Ahn, Hyeonmi;Park, Danbee
    • The Journal of Industrial Distribution & Business
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    • v.10 no.1
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    • pp.69-76
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    • 2019
  • Purpose - The purpose of this paper is to empirically investigate the effectiveness of Offcial Development Assistance (ODA) in recipient countries' economy. ODA is designed to mitigate poverty and stimulate economic growth in the developing countries. We classify total ODA into bilateral ODA and multilateral ODA depending on the number of donor countries. If the ODA flows from one donor country to one recipient country, it is classified as bilateral ODA. If the multiple countries simultaneously become donor countries through the international organizations such as United Nations and World Bank, it is classified as multilateral ODA. This paper compares the effect of bilateral ODA and multilateral ODA in determining recipient countries' economic development, and tries to provide policy implications to Korean ODA. Research design, data, and methodology - Our primary explanatory variables are bilateral and multilateral ODA. Private credit in recipient countries is adopted as additional explanatory variables to capture the level of financial development in recipient countries. We measure the ODA effectiveness using economic growth and quality of life of the recipient countries as the dependent variable. We collect 142 recipient countries' data from OECD statistics, during the period from 1970-2014. Panel least squares estimation with country fixed effect is employed as the empirical model. Results - Our results support that ODA variable has a negatively significant impact on recipient countries' economic growth, while it is positively correlated with human development index. Recipient countries' private credit is positively correlated with economic growth and human development index. The interaction variable of ODA and financial development turns out to be significant in general. We find that the positive effect of ODA depends on recipient countries' financial market development and this effect is stronger in multilateral aid than bilateral one. Conclusions - From the analysis, we have confirmed that the recipient countries financial development is the necessity condition to achieve positive effect of ODA. Based on these results, we suggest that Korean government should increase the share of multilateral funding and pay attention to recipient countries' financial market development to maximize the effectiveness of ODA.

Phenotypic characterization of Hanwoo (native Korean cattle) cloned from somatic cells of a single adult

  • Yang, Byoung-Chul;Lee, Seung-Hwan;Hwang, Seong-Soo;Lee, Hwi-Cheul;Im, Gi-Sun;Kim, Dong-Hoon;Lee, Dong-Kyeong;Lee, Kyung-Tai;Jeon, Ik-Soo;Oh, Sung-Jong;Park, Soo-Bong
    • BMB Reports
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    • v.45 no.1
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    • pp.38-43
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    • 2012
  • We investigated phenotypic differences in Hanwoo cattle cloned from somatic cells of a single adult. Ten genetically identical Hanwoo were generated by somatic cell nuclear transfer from a single adult. Weights at birth, growing pattern, horn and noseprint patterns were characterized to investigate phenotypic differences. The weights of clones at 6 and 12 months were slightly heavier than that of the donor. A horn pattern analysis revealed that seven clones had exactly the same horn pattern as the donor cow, whereas three were different. Although similarities such as general appearance can often be used to identify individual cloned animals, no study has characterized noseprint patterns for this end. A noseprint pattern analysis of all surviving clones showed that all eight animals had distinct noseprints. Four were similar to the donor, and the remaining four had more secondary-like characteristics.