• Title/Summary/Keyword: Cell Fusion

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The role of mitochondria in apoptosis

  • Jeong, Seon-Yong;Seol, Dai-Wu
    • BMB Reports
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    • v.41 no.1
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    • pp.11-22
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    • 2008
  • Apoptosis (programmed cell death) is a cellular self-destruction mechanism that is essential for a variety of biological events, such as developmental sculpturing, tissue homeostasis, and the removal of unwanted cells. Mitochondria play a crucial role in regulating cell death. $Ca^{2+}$ has long been recognized as a participant in apoptotic pathways. Mitochondria are known to modulate and synchronize $Ca^{2+}$ signaling. Massive accumulation of $Ca^{2+}$ in the mitochondria leads to apoptosis. The $Ca^{2+}$ dynamics of ER and mitochondria appear to be modulated by the Bcl-2 family proteins, key factors involved in apoptosis. The number and morphology of mitochondria are precisely controlled through mitochondrial fusion and fission process by numerous mitochondria-shaping proteins. Mitochondrial fission accompanies apoptotic cell death and appears to be important for progression of the apoptotic pathway. Here, we highlight and discuss the role of mitochondrial calcium handling and mitochondrial fusion and fission machinery in apoptosis.

Effect of the Electric Field on the Plant Protoplasts During Cell Fusion (세포융합시 전계하에서 식물세포가 받는 영향에 관한연구)

  • Lee, Sang-Hoon;Lee, Yon-Min;Cha, Hyeon-Cheol
    • Journal of Biomedical Engineering Research
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    • v.17 no.2
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    • pp.173-178
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    • 1996
  • The objective of this paper is to investigate the effect of AC field on the protoplast of plant cells. The results of investigation will be the basis for the development of etectric cell fusion device. For the experiment, we made the electrode and AC and DC pulse generator and observed the behavior of the protoplasts through the inverted microscope which is connected to the monitor and video recorder by the CCD camera. As a result, the numbers of rotating, moving and destructed protoplasts and viability of the protoplasts have close relation to the amplitude of AC field, while the rotation rate is closely related to the frequency of AC pulse.

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Adhesion and Proliferation Behavior of Retinal Pigment Epithelial Cells on Hesperidin/PLGA Films (헤스페리딘/PLGA 필름에서 망막색소상피세포의 부착과 증식거동)

  • Lee, So Jin;Kang, Su Ji;Kim, Hye Yun;Lee, Jung Hwan;Kim, Eun Young;Kwon, Soon Yong;Chung, Jin Wha;Joo, Choun-Ki;Khang, Gilson
    • Polymer(Korea)
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    • v.38 no.1
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    • pp.24-30
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    • 2014
  • Retinal pigment epithelium (RPE) plays an important role in maintaining the visual function and the degeneration of the RPE causes several retinal degeneration disease. In order to fabricate the suitable carrier for RPE transplantation, the hybrid poly(lactide-co-glycolide) (PLGA) film with hesperidin was prepared. Hesperidin has an anti-inflammatory and antioxidant characteristics. ARPE-19 was seeded on hesperidin/PLGA film and then, cell proliferation was determined by the MTT assay, and cell adhesion and cell morphology were confirmed by SEM. Also, RT-PCR was performed to confirm the expression of the specific genes, and AEC immunohistochemical staining was performed to determine the expression of RPE65. As a result, we confirmed that attachment, proliferation and phenotype maintenance of RPE cells were more excellent on hesperidin/PLGA film than PLGA film, thereby we were able to confirm the potential applications of hesperidin/PLGA film as tissue engineering carrier for regeneration of retina.

Effects of Ultraviolet Irradiation on the Differentiation of Cultured Chicken Pectoralis Muscle Cells (培養 鷄胚 筋細胞分化에 미치는 紫外線의 영향)

  • Chung, Hae-Moon;Nham, Sang-Uk
    • The Korean Journal of Zoology
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    • v.24 no.4
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    • pp.189-200
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    • 1981
  • Drastic alterations in myogenesis could be induced by ultraviolet irradiation of the myogenic cells derived from 12 day old chick embryo skeletal muscle. The effects of irradiation on various aspects, including cell division, transformation to myotubes, and morphology of myoblasts and myotubes, were examined. Irradiated cells were smaller in size, and only few cells transformed resulting in smaller size of myotubes with a narrow width. Both the inhibiting actions to cell division and to fusion were more striking when irradiated at earlier stages after plating. As well, cell division and fusion were inhibited more effectively with increasing UV dose and excessive amount caused cell death. A lowering cell density was thought to account for the decrease in myogenesis and possible reasons for the decrease in the capacity for fusion were discussed in view of the results presented in this report and of the findings from other laboratories.

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Effect of Electrical Preactivation of Recipient Cytoplasm on Nuclear Remodelling in Nuclear Transplant Rabbit Embryos (수핵란의 전 활성화가 토끼 핵이식 수정란의 핵 재구성에 미치는 효과)

  • 전병균;김윤연;정기화;곽대오;이효종;최상용;박충생
    • Korean Journal of Animal Reproduction
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    • v.21 no.3
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    • pp.229-238
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    • 1997
  • Chromosome condensation and swelling of the donor nucleus have been known as the early morphological indicators of chromatin remodelling after injection of a foreign nucleus into an enucleated recipient cytoplasm. The effects of non-preactivation and electrical preactivation of recipient cytoplasm, prior to fusing a donor nucleus, on the profile of nuclear remodelling in the nuclear transplant rabbit embryos were evaluated. The embryos of 16-cell stage were collected and synchronized to G1 phase of 32-cell stage. The recipient cytoplasms were obtained by removing the first polar body and chromosome mass by non-disruptive microsurgical procedure. The separated G1 phase blastomeres of 32-cell stage were injected into non-preactivated recipient cytoplasms. Otherwise, the enucleated recipient cytoplasms were preactivated by electrical stimulation and the separated G1 phase blastomeres of 32-cell stage were injected. After culture until 20h post-hCG injection, the nuclear transplant oocytes were electrofused by electrical stimulation. The nuclei of nuclear transplant embryos fused into non-preactivated and/or preactivated recipient cytoplasm were stained by Hoechst 33342 at 0, 1.5, 2, 4, 6, 8, 10 hrs post-fusion and were observed under an fluorescence microscopy. Accurate measurements of nuclear diameter were revealed with an ocular micrometer at 200$\times$. Upon blastomere fusion into non-preactivated recipient cytoplasm, a prematurely chromosome condensation at 1.5 hrs post-fusion and nuclear swelling at 8 hrs post-fusion were occurred as 91.6% and 86.1%, respectively. But the nuclei of nuclear transplant embryos fused into preactivated recipient cytoplasm, as o, pp.sed to non-preactivated recipient cytoplasm, were not occurred chromosome condensation and extensive nuclear swelling. Nuclear diameter fused into non-preactivated and preactivated recipient cytoplasm at hrs post-fusion was 30.2$\pm$0.74 and 15.2$\pm$1.32${\mu}{\textrm}{m}$, respectively. These results indicated that onset of unclear condensation and swelling which was associated with oocytes activation were critical steps in the process of chromatin swelling. Futhermore, complete reprogramming seemed only possible after remodelling of the donor nucleus by chromosome condensation and nuclear swelling.

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Fate of Donor Centrosome and Microtubule Dynamics of Porcine Somatic Cell Nuclear Transfer Embryos

  • Kwon, Dae-Jin;Park, Choon-Keun;Yang, Boo-Keun;Cheong, Hee-Tae
    • Reproductive and Developmental Biology
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    • v.34 no.2
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    • pp.73-79
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    • 2010
  • We investigated the microtubule dynamics, including the inheritance of donor centrosomes and the mitotic spindle assembly occurring during the first mitosis of somatic cell nuclear transfer (SCNT) embryos in pigs. SCNT embryos were fixed 15 min and 1 h after fusion in order to assess the inheritance pattern of the donor centrosome. The distribution and dynamic of the centrosome and microtubule during the first mitotic phase of SCNT embryos were also evaluated. The frequency of embryos evidencing $\gamma$-tubulin spots (centrosome) was 93.2% in the SCNT embryos 15 min after fusion. In the majority of the SCNT embryos (61.5%), however, no centrosome was observed 1 h after fusion. The frequency of the embryos with no or abnormal mitotic spindles 20 h after fusion was 19.6%. The $\gamma$-tubulin spots were detected near the nuclei of somatic cells regardless of cell cycle phase, whereas $\gamma$-tubulin spots in the SCNT embryos were observed only during the inter-anaphase transition. These results showed that the donor centrosome is inherited into the SCNT embryos, but failed to assemble the normal mitotic spindles during first mitotic phase in some SCNT embryos.

Optimum Conditions of pH and Ca2+ Concentration for Electrofusion of Tobacco Protoplasts (담배 워형질체의 전기융합을 위하 pH 및 Ca2+ 농도 최적조건 설정)

  • 오인숙;소상섭;김환규
    • KSBB Journal
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    • v.13 no.4
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    • pp.399-403
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    • 1998
  • This study was carried out to optimize the concentration of Ca2+ and pH of fusion medium which affected electrofusion frequency of protoplasts isolated from Nicotiana tabacum L. (cv. BY4) mesophyll cells and callus. The protoplasts were electrofused in the fusion media containing two different Ca2+ concentrations and three different pH regions. Fusion frequency was lower in the fusion medium containing only 13% mannitol as osmotic stabilizer. However, higher degree of fusion frequency (47.3%) was observed in the fusion medium containing 50mM CaCl2 at pH 10.5 than any other conditions. Cell viability was decreased by Ca2+ and high pH treatment in the fusion media, while fusion frequency was increased. It is concluded that Ca2+ is involved in electrofusion of protoplasts.

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Culture and Fusion of Protoplasts from Potato (Solanum tuberosum L.) and Tobacco (Nicotiana tabacum L.) (감자(Solanum tuberosum L.)와 담배 (Nicotiana tabacum L.)의 원형질체 배양 및 융합)

  • 정상호
    • Journal of Plant Biology
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    • v.30 no.4
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    • pp.287-298
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    • 1987
  • The regenerative capacities of protoplasts isolated from potato (Solamum tuberosum L.) tubers and tobacco (Nicotiana tabacum L.) mesophyll tissues were examined, and then their intergeneric protoplast fusion was carried out. The potato tuber-derived protoplasts proliferated into the calli some of which showed rudimentary shoot-like structures, which had not been attempted before from tubers, while the tobacco protoplasts were regenerated into the whole plants. Intergeneric protoplast fusion between potato and tobacco was carried out and the heteroplasmic fusion products were formed. The first cell division of some of them was observed after 5 days of culture.

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Effect of $Ca^{2+}$ Concentration in Fusion Medium on the Fusion, Nuclear Morphology and Development of Bovine Somatic Cell Nuclear Transfer Embryos (세포 융합액 중의$Ca^{2+}$ 농도가 소 체세포 핵이식란의 융합, 핵형 및 체외발육에 미치는 영향)

  • 조재원;김정익;박춘근;양부근;정희태
    • Korean Journal of Animal Reproduction
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    • v.26 no.1
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    • pp.33-39
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    • 2002
  • This study was conducted to investigate the effect of $Ca^{2+}$ concentration in fusion medium on the fusion, nuclear morphology and the development of bovine somatic cell nuclear transfer embryos. Bovine skin cells were transferred into an enucleated oocyte and fused with cytoplasm in the fusion medium containing with 0.05 to 1.0 mM Cacl$_2$. Nuclear transfer embryos were activated with a combination of A23187 and cycloheximide. Nuclear transfer embryos were fixed at 3 h after fusion or cultured for 7 ~8 days. Fusion rate was significantly (P<0.01) increased by increasing the $Ca^{2+}$ concentrations in the fusion medium from 0.05 mM (56.6%) to 0.5 mM (50.1%) and 1.0 mM (84.3%). More than 80% of reconstituted embryos underwent premature chromosome condensation (PCC) with 0.05, 0.1 mM CaCl$_2$, whereas 54.5% and 59.3% of embryos formed pronucleus (PN) directly without PCC in the 0.5 and 1.0 mM CaCl$_2$, groups. Blastocyst formation rates were significantly (P<0.05) different between 0.1 mM and 1.0 mM CaCl$_2$groups. From the present result, it is suggested that the elevated $Ca^{2+}$ concentrations in fusion medium can enhance the fusion and blastocyst formation rates of bovine nuclear transfer embryos.bryos.