• Title/Summary/Keyword: donor cell

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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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Developmental Efficiency of Bovine Embryos Cloned with Fetal Fibroblast Arrested at G0/G1 Phase

  • Cho, S.R.;Son, W.J.;Park, C.S.;Park, G.J.;S.Y. Choe
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.140-140
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    • 2003
  • The study evaluated the effect of donor cell treatments for G0/Gl synchronization and the donor ceil type on development and incidence of apoptosis in cloned cattle embryos. Primary cultures were established from a female fetus on day 50 of gestation and adult ear skin biopsies. Cells were randomly allocated into 3 experimental treatment groups after 6~8 passages. Group 1 (Confluent), cells were cultured in DMEM supplemented with 10% FBS until 90% confluent. Group 2 (Serum-starvation), cells were cultured in DMEM Supplemented With 0.5% FBS for 5 days. Group 3 (Roscovitine), Cells were cultured in DMEM supplemented with 10% FBS and 30 $\mu$M Roscovitine for 12 h. Cell cycle and apoptosis were analyzed using flow cytometry after labelling with DAPI and YO-PRO-1. At 19 h post-maturation (hpm), enucleated oocytes were reconstructed with donor cells and fused by a single DC pulse (1.6 kV/cm, 60 $\mu$sec). (중략)

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Effects of Donor Somatic Cell Conditions on In Vitro Development of Nuclear Transplanted Porcine Embryos (돼지 공여세포의 조건이 핵이식 수정란의 체외발달에 미치는 영향)

  • 홍승표;박준규;이명열;이지삼;정장용
    • Journal of Embryo Transfer
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    • v.16 no.3
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    • pp.213-221
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    • 2001
  • This study was conducted to examine in vitro developmental ability of porcine embryos after somatic cell nuclear transfer. The porcine ear fell was cultured in vitro for confluency in serum-starvation condition(TCM-199 + 0.5% FBS) far 3~6 days of cell confluency. The zona pellucida of IVM oocytes were partially drilled using laser system. Single somatic cell was individually transferred into enucleated oocytes. And the reconstructed embryos were electrically fused(single DC 1.9kv/cm, 30$\mu$ sec) with 0.3M mannitol. After electrofusion, embryos were activated(single AC 5v/mm, 5sec) and cultured in HCSU-23 medium containing 10% FBS at 39$^{\circ}C$, 5% $CO_2$ in air for 6 to 8 days. The fusion rate of donor cells was 45.6, 36.8 and 46.1% in 3~4, 5~6 days of serum starvation and non serum starvation(N-S), and were 52.7. 53.0 and 51.7% in 1~2. 5~6 and 13~14 passages of donor cell culture, respectively. No significant difference was found in the fusion rate of donor cells by the duration of serum starvation treatment or the number of donor cell passages. By the size of donor cells, however, the fusion rate was significantly higher(P<0.05) for reconstructed embryos derived from 25r $\mu$m $\geq$ site of donor cells (65.3%) than that of 25~30$\mu$ m(42.5%) or 30$\mu$ m(45.5%)$\leq$ cells. The cleavage rate was significantly (P<0.05) higher in 3~4 darts of serum starvation treatment(67.1%) than that in N-S (50.7%) or 5~6 days of starvation(57.1%). The activation rate by the size of donor cells in fused oocytes was 56.5, 68.8 and 58.5%, respectively, and was not significant.

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Cats Cloned from Fetal Fibroblast Cells by Nuclear Transfer

  • Yin, X.J.;Lee, H.S.;Lee, Y.H.;Hwang, W.S.;Kong, I.K.
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2004.10a
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    • pp.26-31
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    • 2004
  • This work was undertaken in order to study the developmental competence of nuclear transfer cat embryo with fetal fibroblast and adult skin fibroblast as donor nuclei. Oocytes wererecovered by mincing the ovaries in Hepes-buffered TCM199 and selected the cumulus oocyte complexes (COCs) with compact cumulus cell mass and dark. Homogenous ooplasm were cultured for maturation in TCM199 + 10% fetal bovine serum (FBS) for 12 hours and used as a source of recipient cytoplast for exogenous somatic nuclei. In Experiment 1, we evaluated the effect donor cell types on the reconstruction and development of cloned embryos. Fusion, first cleavage and blastocyst developmental rate was not different between fetal fibroblast and adult skin cell (71.2 vs. 66.8; 71.0 vs. 57.6; 4.0 vs. 6.1 %, P<0.05). In Experiment 2, cloned embryos were surgically transferred into the oviducts of recipient queens. One of seven recipient queens was delivered naturally 2healthy cloned cats and 1 stillborn from fetal fibroblast cell of male origin after 65 days embryo transfer. One of three recipient queens was delivered naturally 1 healthy cloned cat from adult skin cell of female after 65 days embryo transfer. The cloned cats showed genotypes identical to the donor cell lines, indicating that adult somatic cells can be used for feline cloning.

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Closure of radial forearm free flap donor-site defect with proportional local full-thickness skin graft: case series study of a new design

  • Han, Yoon-Sic;Lee, Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.47 no.6
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    • pp.427-431
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    • 2021
  • Objectives: The aim of this study was to describe a simple, convenient, and reliable new technique using local full-thickness skin graft (FTSG) for skin coverage of a donor-site defect of the radial forearm free flap (RFFF). Patients and Methods: Between April 2016 and April 2021, five patients with oral squamous cell carcinoma underwent mass resection combined with RFFF reconstruction. After RFFF harvesting, donor-site defects were restored by proportional local FTSG. Results: The donor-site defects ranged in size from 24 to 41.25 cm2, with a mean of 33.05 cm2. Good or acceptable esthetic outcomes were obtained in all five patients. There was no dehiscence, skin necrosis, wound infection, or severe scarring at the graft site through the end of the postoperative follow-up period, and no patient had any specific functional complaint. Conclusion: The proportional local FTSG showed promising results for skin coverage of the donor-site defect of the RFFF. This technique could decrease the need for skin grafts from other sites.

Experience of ReCell in Skin Cancer Reconstruction

  • Gilleard, Onur;Segaren, Nicholas;Healy, Ciaran
    • Archives of Plastic Surgery
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    • v.40 no.5
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    • pp.627-629
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    • 2013
  • The ReCell system (Avita Medical) is a cell culture product that allows the immediate processing of a small split-thickness skin biopsy to produce a complete population of cells including keratinocytes, melanocytes, Langerhans cells and fibroblasts. This series is the first to highlight the reconstructive applications of ReCell following ablative skin cancer surgery. The ReCell system was utilized for three patients following skin cancer excision. In two cases, the cells were applied to forehead flap donor sites following nasal reconstruction. In one case, the cells were applied to the calvarial periosteum following wide local excision of a melanoma scar. Assessment of the treated area was performed using the patient and observer scar assessment scale after 1 year. The Patient and Observer Scar Assessment Scale (POSAS) scores for the 2 patients treated with ReCell following forehead flap surgery were 22 and 32. The score for the patient that underwent wide local excision of a melanoma scar was 45. The absence of a donor site, accelerated healing and the satisfactory aesthetic appearance of the mature scars in this series suggest that ReCell may play a useful role in reconstruction following skin cancer excision.

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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Ectopic Overexpression of COTE1 Promotes Cellular Invasion of Hepatocellular Carcinoma

  • Zhang, Hai;Huang, Chang-Jun;Tian, Yuan;Wang, Yu-Ping;Han, Ze-Guang;Li, Xiang-Cheng
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5799-5804
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    • 2012
  • Family with sequence similarity 189, member B (FAM189B), alias COTE1, a putative oncogene selected by microarray, for the first time was here found to be significantly up-regulated in hepatocellular carcinoma (HCC) specimens and HCC cell lines. mRNA expression of COTE1 in HCC samples and cell lines was detected by reverse transcription-polymerase chain reaction (RT-PCR) and real-time PCR, while protein expression of COTE1 in HCC tissues was assessed by immunohistochemistry. In addition, invasion of HCC cells was observed after overexpressing or silencing COTE1. In the total of 48 paired HCC specimens, compared with the adjacent non-cancer tissues, the expression of COTE1 was up-regulated in 31 (p<0.01). In HCC cell lines, COTE1 expression was significantly higher than in normal human adult liver (p<0.01). Overexpression of COTE1 enhanced HCC-derived LM6 and MHCC-L cellular invasion in vitro. In contrast, COTE1 knockdown via RNAi markedly suppressed these phenotypes, as documented in LM3 and MHCC-H HCC cells. Mechanistic analyses indicated that COTE1 could physically associate with WW domain oxidoreductase (WWOX), a tumor suppressor. COTE1 may be closely correlated with invasion of hepatocellular carcinoma (HCC) cells and thus may serve as an effective target for gene therapy.

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.

Performance Comparison of CuPc, Tetracene, Pentacene-based Photovoltaic Cells with PIN Structures

  • Hwang, Jong-Won;Kang, Yong-Su;Park, Seong-Hui;Lee, Hye-Hyun;Jo, Young-Ran;Choe, Young-Son
    • Proceedings of the Korean Vacuum Society Conference
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    • 2010.08a
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    • pp.311-312
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    • 2010
  • The fabricated photovoltaic cells based on PIN heterojunctions, in this study, have a structure of ITO/poly(3, 4-ethylenedioxythiophene)-poly(styrenesulfonate)(PEDOT:PSS)/donor/donor:C60(10nm)/C60(35nm)/2, 9-dimethyl-4, 7-diphenyl-1, 10-phenanthroline(8nm)/Al(100nm). The thicknesses of an active layer(donor:C60), an electron transport layer(C60), and hole/exciton blocking layer(BCP) were fixed in the organic photovoltaic cells. We investigated the performance characteristics of the PIN organic photovoltaic cells with copper phthalocyanine(CuPc), tetracene and pentacene as a hole transport layer. Discussion on the photovoltaic cells with CuPc, tetracene and pentacene as a hole transport layer is focussed on the dependency of the power conversion efficiency on the deposition rate and thickness of hole transport layer. The device performance characteristics are elucidated from open-circuit-voltage(Voc), short-circuit-current(Jsc), fill factor(FF), and power conversion efficiency($\eta$). As the deposition rate of donor is reduced, the power conversion efficiency is enhanced by increased short-circuit-current(Jsc). The CuPc-based PIN photovoltaic cell has the limited dependency of power conversion efficiency on the thickness of hole transport layer because of relatively short exciton diffusion length. The photovoltaic cell using tetracene as a hole transport layer, which has relatively long diffusion length, has low efficiency. The maximum power conversion efficiencies of CuPc, tetracene, and pentacene-based photovoltaic cells with optimized deposition rate and thickness of hole transport layer have been achieved to 1.63%, 1.33% and 2.15%, respectively. The photovoltaic cell using pentacene as a hole transport layer showed the highest efficiency because of dramatically enhanced Jsc due to long diffusion length and strong thickness dependence.

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