• Title/Summary/Keyword: cell aggregation

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Developmental potential of aggregated mouse embryos and production of chimeras after freezing (동결보존한 마우스 집합배의 생존성과 chimera의 생산에 관한 연구)

  • Shin, Sang-tae;Jo, Choong-ho
    • Korean Journal of Veterinary Research
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    • v.30 no.2
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    • pp.231-241
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    • 1990
  • The present study was carried out to investigate the viability of frozen-thawed aggregated mouse embryos and to produce the chimeras. Different phenotypic embryos were obtained by mating ICR female mice with ICR or CBA male mice. The aggregated morulae were produced following aggregation of embryos at 4-, 8- and 12- to 16-cell stage. The desirable stage for the aggregation of the mouse embryos was 8- to 16-cell stage. The post-thawed in vitro survival rates of aggregated embryos in glycerol, DMSO and ethylene glycerol were 51.5, 78.6 and 69.4%, respectively. Although the higher survival was obtained in DMSO, there were no significant differences in the survival rates among the three cryoprotectants. A total of 155 frozen-thawed aggregated embryos were transferred to 11 recipient mice, 3 out of 7 offsprings were born to overt chimera. These experiments have proven that mouse chimeras can be obtained from frozen-thawed aggregated embryos.

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Studies on the Aggregation of H-Y Antibody-Sexed and Bisected Rabbit Embryo (H-Y항체에 의한 토끼배의 성 감별과 이등분 절단 토끼배의 융합에 관한 연구)

  • 최화식;임경순;진동일
    • Korean Journal of Animal Reproduction
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    • v.21 no.2
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    • pp.85-93
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    • 1997
  • These experiments were carried out to examine the development capacity of sexed and then bisected embryo from 8-cell to morula stage. Antisera to histocompatibility-Y(H-Y) antigen were prepared in inbred SD female rat by repeated immunization of spleen cell or testis supernatant from males of same strain. Male and female embryos were separated by delaying development of embryos against H-Y antibody. After sexing, rabbit embryos were bisected and aggregated. The results obtained from the these experiments were summuerized as follows: 1. When mouse and rabbit 8-, 16-cell and morular embryos were culature in H-Y antiserum, the ratio of embryo which has developed to hatching blastocyst was 53.4, 46.3 and 57.4% in mouse embryos, and 49.0, 52.0 and 61.0% in rabbit embryo, respectively. The ratio of mouse and rabbit embryos developed to hatching blastocyst showed nearly natural sex rate(50%), except rabbit mourla showed a little higher ratio(61.0%) as compared to natural sex ratio. 2. When rabbit demi-embryos from 8-, 16-cell embryo and morula were cultured, the percentage of demi-embryos was 70, 68 and 58% without zona pellucida removed, and 62, 69 and 51% with zona pellucida. The rate of aggregation was higher in 8- and 16-cell demi-embryos than in morula demi-embryo. 4. When sexed-demi-embryo was aggregated with another demi-embryo with demi-embryo with same sex, the rate of embryo developed to blastocyst was 60, 50 and 25%, respectively. Eight-cell demi-embryo showed highest rate. In conclusion, it showed that H-Y antiserum which was made by rat spleen cell enabled sexing rabbit embryos. And when rabbit sexed 8-, 16-cell and morula demi-embryo were aggregated, they were developed to eu-blastocyst which suggested the potential of sexed embryo manipulation.

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Numerical Simulation of Blood Cell Motion in a Simple Shear Flow

  • Choi, Choeng-Ryul;Kim, Chang-Nyung;Hong, Tae-Hyub
    • Proceedings of the KSME Conference
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    • 2008.11a
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    • pp.1487-1491
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    • 2008
  • Detailed knowledge on the motion of blood cells flowing in micro-channels under simple shear flow and the influence of blood flow is essential to provide a better understanding on the blood rheological properties and blood cell aggregation. The microscopic behavior of red blood cell (RBCs) is numerically investigated using a fluid-structure interaction (FSI) method based on the Arbitrary-Lagrangian-Eulerian (ALE) approach and the dynamic mesh method (smoothing and remeshing) in FLUENT (ANSYS Inc., USA). The employed FSI method could be applied to the motions and deformations of a single blood cell and multiple blood cells, and the primary thrombogenesis caused by platelet aggregation. It is expected that, combined with a sophisticated large-scale computational technique, the simulation method will be useful for understanding the overall properties of blood flow from blood cellular level (microscopic) to the resulting rheological properties of blood as a mass (macroscopic).

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Inhibition of ${\beta}-amyloid_{1-40}$ Peptide Aggregation and Neurotoxicity by Citrate

  • Park, Yong-Hoon;Kim, Young-Jin;Son, Il-Hong;Yang, Hyun-Duk
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.4
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    • pp.273-279
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    • 2009
  • The accumulation of ${\beta}$-amyloid (A${\beta}$) aggregates is a characteristic of Alzheimer's disease (AD). Furthermore, these aggregates have neurotoxic effects on cells, and thus, molecules that inhibit A${\beta}$ aggregate formation could be valuable therapeutics for AD. It is well known that aggregation of A${\beta}$ depends on its hydrophobicity, and thus, in order to increase the hydrophilicity of A${\beta}$, we considered using citrate, an anionic surfactant with three carboxylic acid groups. We hypothesized that citrate could reduce hydrophobicity and increase hydrophilicity of A${\beta}_{1-40}$ molecules via hydrophilic/electrostatic interactions. We found that citrate significantly inhibited A${\beta}_{1-40}$ aggregation and significantly protected SH-SY5Y cell line against A${\beta}_{1-40}$ aggregates-induced neurotoxicity. In details, we examined the effects of citrate on A${\beta}_{1-40}$ aggregation and on A${\beta}_{1-40}$ aggregates-induced cytotoxicity, cell viability, and apoptosis. Th-T assays showed that citrate significantly inhibited A${\beta}_{1-40}$ aggregation in a concentration-dependent manner (Th-T intensity: from 91.3% in 0.01 mM citrate to 82.1% in 1.0 mM citrate vs. 100.0% in A${\beta}_{1-40}$ alone). In cytotoxicity and viability assays, citrate reduced the toxicity of A${\beta}_{1-40}$ in a concentration-dependent manner, in which the cytotoxicity decreased from 107.5 to 102.3% as compared with A${\beta}_{1-40}$ aggregates alone treated cells (127.3%) and the cell viability increased from 84.6 to 93.8% as compared with the A${\beta}_{1-40}$ aggregates alone treated cells (65.3%). Furthermore, Hoechst 33342 staining showed that citrate (1.0 mM) suppressed A${\beta}_{1-40}$ aggregates-induced apoptosis in the cells. This study suggests that citrate can inhibit A${\beta}_{1-40}$ aggregation and protect neurons from the apoptotic effects of A${\beta}_{1-40}$ aggregates. Accordingly, our findings suggest that citrate administration should be viewed as a novel neuroprotective strategy for AD.

Transient microfluidic approach to the investigation of erythrocyte aggregation: comparison and validation of the method

  • Hou, Jian-Xun;Shin, Se-Hyun
    • Korea-Australia Rheology Journal
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    • v.20 no.4
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    • pp.253-260
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    • 2008
  • A method based on transient shear flow dynamics of red cell aggregates was developed to investigate reversible re-aggregation processes with decreasing shear flow. In the microchannel-flow aggregometry, the aggregated red blood cells that are subjected to continuously decreasing shear stress in microchannel flow were measured with the use of a laser-scattering technique. Both the laser-backscattered intensity and pressure were simultaneously measured with respect to time, resulting in shear stress ranging from $0{\sim}35\;Pa$ for a time period of less than 30 seconds. The time dependent recording of the backscattered light intensity (syllectogram) yielded an upward convex curve with a peak point, which reflected the transition threshold of aggregation in the RBC suspensions. Critical-time and critical-shear stress corresponding to the peak point were examined by varying the initial pressure-differential and the micro channel depth, and these results showed good potential for being used as new aggregation indices. In the present study, these newly proposed indices were also validated by differentiating the effect of fibrinogen on RBC aggregation and then these indices were compared to the conventional indices that were measured by a rotational aggregometer.

Role of post-translational modifications on the alpha-synuclein aggregation-related pathogenesis of Parkinson's disease

  • Yoo, Hajung;Lee, Jeongmin;Kim, Bokwang;Moon, Heechang;Jeong, Huisu;Lee, Kyungmi;Song, Woo Jeung;Hur, Junho K.;Oh, Yohan
    • BMB Reports
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    • v.55 no.7
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    • pp.323-335
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    • 2022
  • Together with neuronal loss, the existence of insoluble inclusions of alpha-synuclein (α-syn) in the brain is widely accepted as a hallmark of synucleinopathies including Parkinson's disease (PD), multiple system atrophy, and dementia with Lewy body. Because the α-syn aggregates are deeply involved in the pathogenesis, there have been many attempts to demonstrate the mechanism of the aggregation and its potential causative factors including post-translational modifications (PTMs). Although no concrete conclusions have been made based on the previous study results, growing evidence suggests that modifications such as phosphorylation and ubiquitination can alter α-syn characteristics to have certain effects on the aggregation process in PD; either facilitating or inhibiting fibrillization. In the present work, we reviewed studies showing the significant impacts of PTMs on α-syn aggregation. Furthermore, the PTMs modulating α-syn aggregation-induced cell death have been discussed.

Involvement of cAMP in the Human Serum-Induced Migration of Adipose-Derived Stem Cells

  • Lee, Minji;Koh, Wonyoung;Kim, Bomee;Chung, Hyeju;Cho, Gahyang;Kim, Haekwon
    • Development and Reproduction
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    • v.20 no.2
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    • pp.101-108
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    • 2016
  • Previously we observed that human adipose-derived stem cells (hADSCs) could form aggregation during culture in the presence of human serum (HS). In the present study, we have examined if the aggregation might result from the cell migration and analyzed the difference of cell adhesivity after culture in various conditions. When cells were cultured in fetal bovine serum (FBS) alone, there was no morphological change. Similarly, cells pretreated with FBS for 1 day or cultured in a mixture of FBS and HS showed little change. In contrast, cells cultured in HS alone exhibited formation of cell-free area (spacing) and/or cell aggregation. When cells cultured in FBS or pretreated with FBS were treated with 0.06% trypsin, almost cells remained attached to the dish surfaces. In contrast, when cells cultured in HS alone were examined, most cells detached from the dish by the same treatment. Treatment of cells with forskolin, isobutylmethyl xanthine (IBMX) or LY294002 inhibited the formation of spacing whereas H89 or Y27632 showed little effect. When these cells were treated with 0.06% trypsin after culture, most cells detached from the dishes as cells cultured in HS alone did. However, cells treated with IBMX exhibited weaker adhesivity than HS alone. Based on these observations, it is suggested that HS treatment might decrease the adhesivity and induce three-dimensional migration of hADSCs, in the latter of which cAMP signaling could be involved.

Numerical Simulation for the Aggregation of Charged Particles (하전입자의 응집성장에 대한 수치적 연구)

  • Park, Hyung-Ho;Kim, Sang-Soo;Chang, Hyuk-Sang
    • Proceedings of the KSME Conference
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    • 2001.06d
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    • pp.605-611
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    • 2001
  • A numerical technique for simulating the aggregation of charged particles was presented with a Brownian dynamic simulation in the free molecular regime. The Langevin equation was used for tracking each particle making up an aggregate. A periodic boundary condition was used for calculation of the aggregation process in each cell with 500 primary particles of 16 nm in diameter. We considered the thermal force and the electrostatic force for the calculation of the particle motion. The morphological shape of aggregates was described in terms of the fractal dimension. The fractal dimension for the uncharged aggregate was $D_{f}=1.761$. The fractal dimension changed slightly for the various amounts of bipolar charge. However, in case of unipolar charge, the fractal dimension decreased from 1.641 to 1.537 with the increase of the average number of charges on the particles from 0.2 to 0.3 in initial states.

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Facile Preparation of Water Dispersible Red Fluorescent Organic Nanoparticles for Cell Imaging

  • Luo, Miao
    • Bulletin of the Korean Chemical Society
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    • v.35 no.6
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    • pp.1732-1736
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    • 2014
  • Red fluorescent organic nanopaticles (FONs) based on a diarylacrylonitrile derivative conjugated molecule were facilely prepared by surfactant modification. Such red FONs showed excellent water solubility and biocompatibility, making them promising for cell imaging applications.

Embryo Aggregation Promotes Derivation Efficiency of Outgrowths from Porcine Blastocysts

  • Lee, Sang-Goo;Park, Jin-Kyu;Choi, Kwang-Hwan;Son, Hye-Young;Lee, Chang-Kyu
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.11
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    • pp.1565-1572
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    • 2015
  • Porcine embryonic stem cells (pESCs) have become an advantageous experimental tool for developing therapeutic applications and producing transgenic animals. However, despite numerous reports of putative pESC lines, deriving validated pESC lines from embryos produced in vitro remains difficult. Here, we report that embryo aggregation was useful for deriving pESCs from in vitro-produced embryos. Blastocysts derived from embryo aggregation formed a larger number of colonies and maintained cell culture stability. Our derived cell lines demonstrated expression of pluripotent markers (alkaline phosphatase, Oct4, Sox2, and Nanog), an ability to form embryoid bodies, and the capacity to differentiate into the three germ layers. A cytogenetic analysis of these cells revealed that all lines derived from aggregated blastocysts had normal female and male karyotypes. These results demonstrate that embryo aggregation could be a useful technique to improve the efficiency of deriving ESCs from in vitro-fertilized pig embryos, studying early development, and deriving pluripotent ESCs in vitro in other mammals.