• Title/Summary/Keyword: EB formation

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Formation of Functional Cardiomyocytes Derived from Mouse Embryonic Stem Cells

  • 신현아;김은영;이영재;이금실;조황윤;박세필;임진호
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.76-76
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    • 2003
  • Pluripotent embryonic stem cells can differentiate into beating cardiomyocytes with proper culture conditions and stimulants via embryo-like aggregates. We describe here the use of mouse embryonic stem (mES03) cells as a reproducible differentiation system for cardiomyocyte. mES03 cells growing in colonies were dissociated and allowed to re-aggregated in suspension [embryoid body (EB) formation〕. To induce cardiomyocytic differentiation, cells were exposed to 0.75% dimethyl sulfoxide (DMSO) during EB formation for 4 days and then another 4 days without DMSO (4+/4-). Thus treated EB was plated onto gelatin-coated dishes for differentiation. Spontaneously contracting colonies which appeared in approximately 4~5 days upon differentiation were mechanically dissected, enzymatically dispersed, plated onto coverslips, and then incubated for another 48~72 hrs. By RT-PCR, robust expression of cardiac myosin heavy chain $\alpha$, cardiac muscle heavy polypeptide 7 $\beta$($\beta$-MHC), cardiac transcription factor GATA4, and skeletal muscle-specific $\alpha$$_1$-subunit of the L-type calcium channel ($\alpha$$_1$CaC $h_{sm}$ ) were detected as early as 8 days after EB formation, but message of cardiac muscle-specific $\alpha$$_1$-subunit of the L-type calcium channel ($\alpha$$_1$CaCh) were reveled at a low level. In contrast, expression of myosin light chain (MLC-2V) and atrial natriuretic factor (ANF) were not detected during EB formation for 8 days. However, a strong expression of the atrial-specific ANF gene was expressed from day 8 onward, which were remained constant in EB. (cardiac specialization and terminal differentiation stage). Electrophysiological examination of spontaneously contracting cells showed ventricle-like action potential 17 days after the EB formation. This study indicates that mES03 cell-derived cardiomyocytes via 4+/4- protocol displayed biochemical and electrophysiological properties of subpopulation of cardiomyocytes.

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DENTAL CARIES CONTROL IN A GIRL WITH RECESSIVE DYSTROPHIC EPIDERMOLYSIS BULLOSA UNDER THE GENERAL ANESTHESIA : A CASE REPORT (열성 이영양성 수포성 표피박리증 환자에서 전신마취를 통한 치과치료: 증례 보고)

  • Cho, Seong-Hyeon;Song, Je-Seon;Lee, Hyo-Seol;Choi, Hyung-Jun;Choi, Byung-Jai;Kim, Seong-Oh;Lee, Jae-Ho
    • The Journal of Korea Assosiation for Disability and Oral Health
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    • v.8 no.2
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    • pp.109-112
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    • 2012
  • Epidermolysis bullosa (EB) is an inherited skin disease characterized by blister formation of the skin following just minor trauma or spontaneously. The main classification of EB is based on the localization of the blistering. In epidermolysis bullosa simplex, the blister formation is intraepidermal; in epidermolysis bullosa junctionalis the blister formation in the epidermal-dermal junction; in the dystrophic forms of EB blister formation is intradermal. Oral manifestations of recessive dystrophic EB are characterized by rampant caries, crowding, microstomia, ankyloglossia, vestibular obliteration. Dental therapy may be well provided to patient with recessive dystrophic EB by using general anesthesia especially in case of poor cooperation. A two years old girl with recessive dystrophic EB visited our clinic for rampant caries. She was hospitalized for severe anemia and fever, and we provided dental therapy under general anesthesia. Extraction of incisors and stainless steel crown restoration of primary molars were performed.

Design and Implementation of an ebXML CPP/CPA Editor (ebXML CPP/CPA 문서 편집기 설계 및 구현)

  • Moonyoung Chun;Soo-Young Oh;HyeonSung Cho;Hyun-Kyu Cho
    • Proceedings of the CALSEC Conference
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    • 2001.08a
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    • pp.241-251
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    • 2001
  • B2C e-commerce is now sufficiently stable. Judging from its success, we can expect B2B to similarly improve business processes for a better return on investment. To achieve B2B e-commerce, many consortia have been trying to develop a B2B framework. As a result, the ebXML is started as an international standard for B2B e- commerce based on XML. The exchange of information between two parties requires each party to know the other party's supported business collaborations, the other party's role in the business collaboration, and the technology details about how the other party sends and receive messages. In some cases, it is necessary for the two parties to reach agreement on some of the details. The ebXML CPP and CPA specification defines how to describe these. In this paper, we'll describe our design and implementation of an ebXML CPP and CPA editor which enables each party to create and edit his profiles and which automatically composites two CPPs. Although it is possible to create and edit CPP and CPA documents with general-purpose XML editor, it is not easy to create and edit CPP and CPA documents with a general-purpose XML editor. Moreover, the detailed procedures for CPA formation from CPPs are currently not provided in the specification. Therefore, we propose two approaches for a CPA formation in addition to easy-to-use user interface.

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Differentiation of Human Embryonic Stem Cells into Germ Cell and Culture Condition for Single Embryonic Stem Cells Dissociated by Enzyme (인간 배아줄기세포의 생식세포로의 분화 및 효소에 의해 분리된 단일줄기세포 배양조건)

  • Chi, Hee-Jun;Choi, Soon-Young;Chung, Da-Yeon
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.1
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    • pp.13-23
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    • 2010
  • Objective: The present study was carried out to induce differentiation of human embryonic stem cells (hESCs) into germ cells and to establish a culture condition for single hESCs dissociated by enzyme. Methods: Embryonic body (EB) was formed by hanging drop culture for 3 days from hESCs colony. The EBs were cultured in the medium supplemented with retionic acid (RA) or/and bone morphogenetic protein-4 (BMP4) for 14 days to differentiate into germ cells. Germ cell specific markers, c-kit and VASA were used for immunohistochemistry of EB. Human ESCs colonies were dissociated into single cells by Collagenase, Tryple and Accutase, and then colony formation rate of the single cells was examined. Rho-associated kinase inhibitor (ROCK inhibitor, Y27632) was added into the culture medium of single cells to reduce the apoptotic damage during the dissociation. Results: Single cells dissociated with Tryple or Accutase showed higher colony formation rates compared to the cells dissociated with Collagenase. Seeding of $5{\times}10^3$ cells/well (4 well dish) was efficient to obtain high colony formation rate compared to other concentrations of seeding cell. Addition of Y27632 significantly increased the colony formation rate of the single cells dissociated by Tryple. Immunohistochemistry of EB with c-kit and VASA markers showed a weak fluorescence signals compared to the signals from the testicular tissue. Conclusion: Dissociation with Tryple was useful to obtain healthy single cells and addition of Y27632 was beneficial for survival and colony formation of the single cells. Unlike other studies, we just observed a dim fluorescence staining of the germ cell markers, probably caused by the short-term culture for the differentiation of EB compared to other studies.

Removal of EDCs from Industrial Sludge by Electron Beam

  • Han, Bumsoo;Kim, Jinkyu;Kim, Yuri;Jung, Seungtae;Park, Junhyung;Choi, Jangseung
    • Journal of Radiation Industry
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    • v.8 no.2
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    • pp.71-76
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    • 2014
  • Endocrine disrupting chemicals (EDCs) and potential EDCs are mostly man-made, found in various materials such as pesticides, additives or contaminants in food, and personal care products. The high energy ionizing radiation has the ability to remove the EDCs with a very high degree of reliability and in a clean and efficient manner. The ionizing radiation interacts with EDCs both directly and indirectly. Direct interaction takes place with EDCs and the structure of EDCs is destroyed or changed. During indirect interaction, radiolysis products of water result in the formation of highly reactive intermediates which then react with the target molecules, culminating in structural changes. To confirm the radiation reduction of EDCs in industrial sludge, a pilot scale experiment up to 50 kGy of electron beam (EB) was conducted with samples from the textile dyeing industries. The experimental result showed the over 90% of reduction of Nonylphenol (NP) and Di(2-ethylhexyl) phthalate (DEHP) at around 10 kGy of absorbed doses.

The Effect of Oxide Formation on the Lifetime of Plasma Sprayed or EB-PVD Thermal Barrier Coatings (플라즈마 용사 및 EB-PVD에 의한 열벽코팅 수명에 대한 산화물 생성의 영향)

  • ;R.D.Sisson;Jr
    • Journal of the Korean institute of surface engineering
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    • v.27 no.2
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    • pp.91-98
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    • 1994
  • For the plasma sprayed as well as the EB-PVD thermal barrier coatings, the fracture paths within the oxidation products developed at the interface between the partially stabilized zirconia ceramic coating and NiCoCrAlY bond coat during cyclic thermal oxidation has been investigated. It was observed that the fracture in the oxidation products primarily took place within the oxide such as $Ni_{1-x}Co_3(Al_,Cr)_2O_4$ or at the interface between the oxide and $Al_2O_3$. It was found that Al2O3 developed first, followed by the Ni/Co/Cr rich oxides such as ,,$Ni_{1-x}Co_x(Al_,Cr)_2O_4$ $Cr_2O_3$and NiO at the interface between the ceramic coating and the bond coat in a cyclic high temperature environment. It was therfore concluded that the formation of the oxide containing Ni, Cr and Co was a life-limiting event for thermal barrier coatings during cyclic thermal oxidation.

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Effects of electron beam irradiation on the superconducting properties of YBCO thin films

  • Lee, Y.J.;Choi, J.H.;Jun, B.H.;Joo, J.;Kim, C.S.;Kim, C.J.
    • Progress in Superconductivity and Cryogenics
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    • v.18 no.4
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    • pp.15-20
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    • 2016
  • The effects of electron beam (EB) irradiation on the superconducting critical temperature ($T_c$) and critical current density ($J_c$) of YBCO films were studied. The YBCO thin films were irradiated using a KAERI EB accelerator with an energy of 0.2 MeV and a dose of $10^{15}-10^{16}e/cm^2$. A small $T_c$ decrease and a broad superconducting transition were observed as the EB dose increased. The value of $J_cs$ (at 20 K, 50 K and 70 K) increased at doses of $7.5{\times}10^{15}$ and $2.2{\times}10^{16}e/cm^2$. However, $J_cs$ decreased as the dose increased further. The X-ray diffraction (XRD) analysis showed that the c axis of YBCO was elongated and the full width at half maximum (FWHM) increased as the dose increased, which is strong evidence of the atomic displacement by EB irradiation. The transmission electron microscopy (TEM) showed that the amorphous layer formed in the vicinity of the surfaces of the irradiated films. The amorphous phase was often present as an isolated form in the interior of the films. In addition to the formation of the amorphous phase, many striations running along the a-b direction of YBCO were observed. The high magnification lattice image showed that the striations were stacking faults. The enhancement of $J_c$ by EB irradiation is likely to be due to the lattice distortion and the formation of defects such as vacancies and stacking faults. The decrease in $J_c$ at a high EB dose is attributed to the extension of the amorphous region of a non-superconducting phase.

Conjugation of vascular endothelial growth factor to poly lactic-co-glycolic acid nanospheres enhances differentiation of embryonic stem cells to lymphatic endothelial cells

  • Yoo, Hyunjin;Choi, Dongyoon;Choi, Youngsok
    • Animal Bioscience
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    • v.34 no.4
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    • pp.533-538
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    • 2021
  • Objective: Pluripotent stem cell-derived lymphatic endothelial cells (LECs) show great promise in their therapeutic application in the field of regenerative medicine related to lymphatic vessels. We tested the approach of forced differentiation of mouse embryonal stem cells into LECs using biodegradable poly lactic-co-glycolic acid (PLGA) nanospheres in conjugation with growth factors (vascular endothelial growth factors [VEGF-A and VEGF-C]). Methods: We evaluated the practical use of heparin-conjugated PLGA nanoparticles (molecular weight ~15,000) in conjugation with VEGF-A/C, embryoid body (EB) formation, and LEC differentiation using immunofluorescence staining followed by quantification and quantitative real-time polymerase chain reaction analysis. Results: We showed that formation and differentiation of EB with VEGF-A/C-conjugated PLGA nanospheres, compared to direct supplementation of VEGF-A/C to the EB differentiation media, greatly improved yield of LYVE1(+) LECs. Our analyses revealed that the enhanced potential of LEC differentiation using VEGF-A/C-conjugated PLGA nanospheres was mediated by elevation of expression of the genes that are important for lymphatic vessel formation. Conclusion: Together, we not only established an improved protocol for LEC differentiation using PLGA nanospheres but also provided a platform technology for the mechanistic study of LEC development in mammals.

Timed Artificial Insemination or Embryo Transfer using CIDR, Estradiol Benzoate and Prostaglandin $F_{2{\alpha}}$ for the Rebreeding of Korean Native Donor Cattle

  • Son, Dong-Soo;Choe, Chang-Yong;Cho, Sang-Rae;Choi, Sun-Ho;Kim, Hyun-Jong;Lee, Je-In;Kim, Ill-Hwa
    • Journal of Embryo Transfer
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    • v.23 no.2
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    • pp.81-86
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    • 2008
  • This study compared the pregnancy rates of Korean native donor cattle after either a timed artificial insemination (TAI) or embryo transfer (TET) following the synchronization of ovulation using a controlled internal drug release (CIDR) device together with estradiol benzoate (EB) and prostaglandin $F_{2{\alpha}}$ ($PGF_{2{\alpha}}$). Fifty five cows and 8 heifers which had been previously used for embryo production were assigned to two treatments: (1) Thirty-two cattle received a CIDR device and 2 mg EB (Day 0), 25 mg $PGF_{2{\alpha}}$ injection at the time of CIDR removal on Day 7, and 1 mg EB injection on Day 8. All of the cattle received a TAI 30 h (Day 9) after the second EB injection (TAI group). (2) Thirty-one cattle received the same hormonal treatments as in the TAI group. The cattle with corpus luteum (CL) received a TET on Day 16 using frozen-thawed embryos (TET group). Ultrasonographic observations demonstrated that the proportion of cattle with synchronized ovulation on Day 10 and the concomitant formation of new CL on Day 13 did not differ between groups (p>0.05); the overall mean rates were 65.1 and 73.0%, respectively. The conception and pregnancy rates did not differ (p>0.05) between the TAI (12.5% and 12.5%) and TET groups (13.0% and 9.7%), respectively. We conclude that the pregnancy rate following TAI or TET in Korean native donor cattle was poor, which might be due in part to a poor synchrony of ovulation and concomitant CL formation.

Functional Cardiomyocytes Formation Derived from Mouse Embryonic Stem Cells

  • Shin, Hyun-Ah;Lee, Keum-Sil;Cho, Hwang-Yoon;Park, Sae-Young;Kim, Eun-Young;Lee, Young-Jae;Park, Se-Pill;Lim, Jin-Ho
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.100-100
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    • 2003
  • Pluripotent embryonic stem (ES) cells differentiate spontaneously into beating cardiomyocytes via embryo-like aggregates. We describe the use of mouse embryonic stem (mES03) cells as a reproducible differentiation system for cardiomyocyte. To induce cardiomyocytic differentiation, mES03 cells were dissociated and allowed to aggregate (EB formation) at the presence of 0 75% dimethyl sulfoxide (DMSO) for 4 days and then another 4 days without DMSO (4+/4-). Thus treated EBs were plated onto gelatin-coated dish for differentiation. Spontaneously contracting colonies which appeared in approximately 4-5 days upon differentiation. Expression of cardiac-specific genes were determined by RT-PCR. Rebust expression of myosin light chain (MLC-2V), cardiac myosin heavy chain $\alpha$, cardiac muscle heavy polypeptide 7 $\beta(\beta$-MHC), cardiac transcription factor GATA4 and skeletal muscle-specific ${\alpha}_1$-subunit of the L-type calcium channel (${\alpha}_1 CaCh_{sm}$) were detected as early as 8 days after EB formation, but message of cardiac muscle-specific $\alpha$$_1$-subunit of the L-type calcium channel (${\alpha}_1$CaCh) were revealed at a low level. Strikingly, the expression of atrial natriuretic factor (ANF) was not detected. When spontaneous contracting cell masses were examined their electrophysiological features by patch-clamp technique, it showed ventricle-like action potential 17 days after the EB formation. This study indicates that mES03 cell-derived cardiomyocytes displayed biochemical and electrophysiological properties of cardiomyocytes and DMSO enhanced development of cardiomyocytes in 4+/4- method.

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