• Molecules and Cells
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• v.46 no.4
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• pp.209-218
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• 2023

In induced pluripotent stem cells (iPSCs), pluripotency is induced artificially by introducing the transcription factors Oct4, Sox2, Klf4, and c-Myc. When a transgene is introduced using a viral vector, the transgene may be integrated into the host genome and cause a mutation and cancer. No integration occurs when an episomal vector is used, but this method has a limitation in that remnants of the virus or vector remain in the cell, which limits the use of such iPSCs in therapeutic applications. Chemical reprogramming, which relies on treatment with small-molecule compounds to induce pluripotency, can overcome this problem. In this method, reprogramming is induced according to the gene expression pattern of extra-embryonic endoderm (XEN) cells, which are used as an intermediate stage in pluripotency induction. Therefore, iPSCs can be induced only from established XEN cells. We induced XEN cells using small molecules that modulate a signaling pathway and affect epigenetic modifications, and devised a culture method which can produce homogeneous XEN cells. At least 4 passages were required to establish morphologically homogeneous chemically induced XEN (CiXEN) cells, whose properties were similar to those of XEN cells, as revealed through cellular and molecular characterization. Chemically iPSCs derived from CiXEN cells showed characteristics similar to those of mouse embryonic stem cells. Our results show that the homogeneity of CiXEN cells is critical for the efficient induction of pluripotency by chemicals.

• Journal of Animal Reproduction and Biotechnology
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• v.38 no.3
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• pp.109-120
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• 2023

Background: Pluripotent stem cells (PSCs) including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) offer the immense therapeutic potential in stem cell-based therapy of degenerative disorders. However, clinical trials of human ESCs cause heavy ethical concerns. With the derivation of iPSCs established by reprogramming from adult somatic cells through the transgenic expression of transcription factors, this problems would be able to overcome. In the present study, we tried to differentiate porcine iPSCs (piPSCs) into endothelial cells (ECs) for stem cell-based therapy of vascular diseases. Methods: piPSCs (OSKMNL) were induced to differentiation into ECs in four differentiation media (APEL-2, APEL-2 + 50 ng/mL of VEGF, EBM-2, EBM-2 + 50 ng/mL of VEGF) on cultured plates coated with matrigel^® (1:40 dilution with DMEM/F-12 medium) for 8 days. Differentiation efficiency of these cells were exanimated using qRT-PCR, Immunocytochemistry, Western blotting and FACS. Results: As results, expressions of pluripotency-associated markers (OCT-3/4, SOX2 and NANOG) were higher observed in all porcine differentiated cells derived from piPSCs (OSKMNL) cultured in four differentiation media than piPSCs as the control, whereas endothelial-associated marker (CD-31) in the differentiated cells was not expressed. Conclusions: It can be seen that piPSCs (OSKMNL) were not suitable to differentiate into ECs in the four differentiation media unlike porcine epiblast stem cells (pEpiSCs). Therefore, it would be required to establish a suitable PSCs for differentiating into ECs for the treatment of cardiovascular diseases.

• Journal of Veterinary Science
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• v.25 no.2
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• pp.31.1-31.8
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• 2024

Background: Recently, there has been a growing interest in stem cells for human medicine. Limited feline endometrial mesenchymal stem cell (fEM-MSC) research in veterinary medicine necessitates reporting for future feline disease research and therapy. Objectives: This study aimed to isolate fEM-MSCs from feline endometrial tissues and evaluate their morphology, proliferative ability, differentiation ability, and immunophenotype. Methods: Feline endometrial tissues were obtained from the ovariohysterectomies of healthy cats and isolated using an enzymatic method. The morphology and proliferative ability of the isolated cells were assessed using a doubling time (DT) assay from passages 3 to 6 (P3 - P6). We measured pluripotency gene expressions of cells in P2 using quantitative real-time polymerase chain reaction (qRT-PCR). To investigate MSC characteristics, a trilineage differentiation assay was conducted in P4, and cells in P4 were immunophenotyped using flow cytometry. Results: fEM-MSCs showed a typical spindle-shaped morphology under a microscope, and the DT was maintained from P3 to P6. fEM-MSCs could differentiate into adipocytes, osteoblasts, and chondrocytes, and expressed three pluripotency markers (OCT4, SOX2, and NANOG) by qRT-PCR. Immunophenotypic analysis showed that the fEM-MSCs were CD14 ^-, CD34 ^-, CD45 ^-, CD9⁺, and CD44⁺. Conclusions: In this study, the feline endometrium was a novel source of MSCs, and to the best of our knowledge, this is the first report on the isolation method and characteristics of fEM-MSCs.

• Development and Reproduction
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• v.12 no.1
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• pp.87-95
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• 2008

Neural stem/precursor derived from pluripotent human embryonic stem cells (hESCs) has considerable therapeutic potential due to their ability to generate various neural cells which can be used in cell-replacement therapies for neurodegenerative diseases. However, production of neural cells from hESCs remains technically very difficult. Understanding neural-tube like rosette characteristic neural precursor cells from hESCs may provide useful information to increase the efficiency of hESC neural differentiation. Generally, neural rosettes were derived from differentiating hEBs in attached culture system, however this is time-consuming and complicated. Here, we examined if neural rosettes could be formed in suspension culture system by bypassing attachment requirement. First, we tested whether the size of hESC clumps affected the formation of human embryonic bodies (hEBs) and neural differentiation. We confirmed that hEBs derived from $500{\times}500\;{\mu}m$ square sized hESC clumps were effectively differentiated into neural lineage than those of the other sizes. To induce the rosette formation, regular size hEBs were derived by incubation of hESC clumps($500{\times}500\;{\mu}m$) in EB medium for 1 wk in a suspended condition on low attachment culture dish and further incubated for additional $1{\sim}2$ wks in neuroectodermal sphere(NES)-culture medium. We observed the neural tube-like rosette structure from hEBs after $7{\sim}10$ days of differentiation. Their identity as a neural precursor cells was assessed by measuring their expressions of neural precursor markers(Vimentin, Nestin, MSI1, MSI2, Prominin-1, Pax6, Sox1, N-cadherin, Otx2, and Tuj1) by RT-PCR and immunofluorescence staining. We also confirmed that neural rosettes could be terminally differentiated into mature neural cell types by additional incubation for $2{\sim}6$ wks with NES medium without growth factors. Neuronal(Tuj1, MAP2, GABA) and glial($S100{\beta}$ and GFAP) markers were highly expressed after $2{\sim}3$ and 4 wks of incubation, respectively. Expression of oligodendrocyte markers O1 and CNPase was significantly increased after $5{\sim}6$ wks of incubation. Our results demonstrate that rosette forming neural precursor cells could be successfully derived from suspension culture system and that will not only help us understand the neural differentiation process of hESCs but also simplify the derivation process of neural precursors from hESCs.

• Journal of the Korean Society of Marine Environment & Safety
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• v.26 no.2
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• pp.163-174
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• 2020

Particulate matter levels are rapidly increasing daily, and this can affect human health. Therefore, air pollutant emissions from sea vessels require management. This study evaluates the status of air pollutants, focusing on air pollutant emissions from the vessels of the Korea Coast Guard (KCG), and proposes national management measures to reduce emissions. According to a report recently released (2018) by the National Institute of Environmental Research (NIER), emissions from vessels constituted 6.4 % of the total domestic emissions, including 13.1 % NO_x, 10.9 % SO_x, and 9.6 % particulate matter (PM10/PM2.5). Among the rates of pollutant emission from vessels, the emission rates of domestic and overseas cargo vessels were the highest (50.6 %); the ratio of fishing boats was 42.6 %. With respect to jurisdictional sea area, 44.1 % of the emissions are from the south sea, including the Busan and Ulsan ports, and 24.8 % of the emissions are from the west sea, including the Gwangyang and Yeosu ports. The KCG inspects boarding lines to manage emission conditions and regulate air pollutant emissions, but it takes time and effort to operate various discharge devices and measure fuel oil standards. In addition, owing to busy ship schedules, inspection documents are limited in terms of management. Therefore, to reduce the air pollutant emissions of such vessels, regulations will be strengthened to check for air pollutants, and a monitoring system based on actual field data using KCG patrol ships will be established, for each sea area, to manage the emissions of such vessels. Furthermore, there is a need for technological development and institutional support for the introduction of environmentally friendly vessels.

• Korean Chemical Engineering Research
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• v.40 no.6
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• pp.746-751
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• 2002

Fluidized bed combustion is a coal combustion technology that can reduce both SOx and NOx emission; SOx is removed by limestone that is fed into the combustion chamber and the NOx is reduced by low temperature combustion in a fluidized bed combustor and air stepping, but $N_2O$ generation is quite high. $N_2O$ is not only a greenhouse gas but also an agent of ozone destruction in the stratosphere. The calcium oxide(CaO) is known to be a catalyst of $N_2O$ decomposition. This study of $N_2O$ decomposition reaction in fixed bed reactor packed over CaO bed has been conducted. Effects of parameters such as concentration of inlet $N_2O$ gas, reaction temperature, CaO bed height and effect of $CO_2$, NO, $O_2$ gas on the decomposition reaction have been investigated. As a result of the experiment, it has been shown that $N_2O$ decomposition reaction increased with the increasing fixed bed temperature. While conversion of the reaction was decreased with increasing $CO_2$ concentration. Also, under the present of NO, the conversion of $N_2O$ decomposition is decreased. From the result of kinetic study gained the heterogeneous reaction rate on $N_2O$ decomposition. In the case of $N_2O$ decomposition over CaO, heterogeneous reaction rate is. $\frac{d[N_2O]}{dt}=\frac{3.86{\times}10^9{\exp}(-15841/R)K_{N_2O}[N_2O]}{(1+K_{N_2O}[N_2O]+K_{CO_2}[CO_2])}$. In this study, it is found that the calcium oxide is a good catalyst of $N_2O$ decomposition.

• Journal of Life Science
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• v.29 no.8
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• pp.895-903
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• 2019

The aim of the present study was to improve the cell viability of human dental papilla derived single-induced pluripotent stem cells (iPSCs) using a Rho-associated protein kinase (ROCK) inhibitor, Y-27632. The iPSCs were produced using an episomal plasmid-based reprogramming method. After cell separation using trypsin, the iPSCs were treated with 0, 0.5, 1, 2.5, 5, 7.5, or $10{\mu}M$ Y-27632 for 5 d. Cell viability increased significantly following the $5{\mu}M$ Y-27632 treatment (p<0.05). When the iPSCs were exposed to medium containing $10{\mu}M$ Y-27632 for 0, 1, 2, 3, 4, and 5 d, the cell viability rate increased significantly in accordance with the cell viability rate (p<0.05). To evaluate the effect of the Y-27632 treatment on stemness characteristics, the expression of stem cell-specific transcripts and telomerase activity were investigated in the iPSCs treated with $10{\mu}M$ Y-27632 for 5 d. The expression levels of stem cell-specific transcripts, such as OCT-4, NONOG, and SOX-2, and telomerase activity were not significantly different in the iPSCs treated with $10{\mu}M$ Y-27632 as compared with those of untreated control iPSCs (p>0.05). Taken together, the results demonstrated that cell viability can be improved by treatment with the ROCK inhibitor Y-27632, without losing iPSC stemness characteristics.

• Journal of the Korean Society of Marine Environment & Safety
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• v.29 no.6
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• pp.697-703
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• 2023

As a part of the global industrial efforts to reduce environmental pollution owing to air pollution, regulations have been established by the International Maritime Organization (IMO). The IMO has implemented various regulations such as EEXI, EEDI, and CII to reduce air pollution emissions from ships. They are also promoting measures to decrease the power consumption in ships, aiming to conserve energy. Most of the power used in ships is consumed by electric motors. Among the motors installed on ships, the engine room blower that takes up a significant load, operates at a constant irrespective of demand. Therefore, energy savings can be expected through frequency control. In this study, we demonstrated the efficacy of energy savings by controlling the frequency of the electric motor of the generator blower that supplies combustion air to the generator's turbocharger. The system was modeled based on the output data of the turboharger outlet temperature in response to the blower frequency inpu. A PI control system was established to control the frequency with the target being the turbocharger outlet temperature. By maintaining the turbocharger design standard outlet temperature and controlling the blower frequency, we achieved an annual energy saving of 15,552kW in power consumption. The effectiveness of energy savings through frequency control of blower fans was verified during the summer (April to September) and winter (March to October) periods. Based on this, we achieved annual fuel cost savings of 6,091 thousand won and reduction of 8.5 tons of carbon dioxide, 2.4 kg of SOx, and 7.8 kg of NOx air pollutants on the training ship.

• Journal of Embryo Transfer
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• v.29 no.1
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• pp.91-99
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• 2014

Oxygen consumption is a useful parameter for evaluating mammalian embryo quality, since individual bovine embryos was noninvasively quantified by scanning electrochemical microscopy (SECM). Recently, several approaches have been used to measure the oxygen consumption rates of individual embryos, but relationship between oxygen consumption and pregnancy rates of Hanwoo following embryo transfer has not yet been reported. In this study, we measured to investigate the correlation between oxygen consumption rate and pregnancy rates of Hanwoo embryo using a SECM. In addition to, the expression of pluripotent gene and anti-oxidant enzyme was determined using real-time PCR by extracting RNA according to the oxygen consumption of in vivo embryo. First, we found that the oxygen consumption significantly increased in blastocyst-stage embryos (blastocyst) compared to early blastocyst stage embryos, indicating that oxygen consumption reflects the embryo quality (Grade I). Oxygen consumption of blastocyst was measured using a SECM and total cell number of in vitro blastocyst was enumerated by counting cells stained by propidium iodide. The oxygen consumption or GI blastocysts were significantly higher than those of GII blastocysts ($10.2{\times}10^{15}/mols^{-1}$ versus $6.4{\times}10^{15}/mols^{-1}$, p<0.05). Total cell numbers of in vitro blastocysts were 74.8, 90.7 and 110.2 in the oxygen consumption of below 10.0, 10.0~12.0 and over $12.0{\sim}10^{15}/mols^{-1}$, respectively. Pregnant rate in recipient cow was 0, 60 and 80% in the transplantation of embryo with the oxygen consumption of below 10.0, 10.0~12.0 and over $12.0{\times}10^{15}/mols^{-1}$, respectively. GPX1 and SOD1 were significantly increased in over -10.0 group than below 10.0 groups but in catalase gene, there was no significant difference. On the other hand, In OCT-4 and Sox2, pluripotent gene, there was a significant difference (p<0.05) between the below-10.0 ($0.98{\pm}0.1$) and over 10.0 ($1.79{\pm}0.2$). In conclusion, these results suggest that measurement of oxygen consumption maybe help increase the pregnant rate of Hanwoo embryos.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.2
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• pp.266-277
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• 2014

Somatic cell nuclear transfer (SCNT) has generally demonstrated that a differentiated cell can convert into a undifferentiated or pluripotent state. In the SCNT experiment, nuclear reprogramming is induced by exposure of introduced donor nuclei to the recipient cytoplasm of matured oocytes. However, because the efficiency of SCNT still remains low, a combination of SCNT technique with the ex-ovo method may improve the normal development of SCNT embryos. Here we hypothesized that treatment of somatic cells with extracts prepared from the germinal vesicle (GV) stage Siberian sturgeon oocytes prior to their use as nuclear donor for SCNT would improve in vitro development. A reversible permeability protocol with $4{\mu}g/mL$ of digitonin for 2 min at $4^{\circ}C$ in order to deliver Siberian sturgeon oocyte extract (SOE) to porcine fetal fibroblasts (PFFs) was carried out. As results, the intensity of H3K9ac staining in PFFs following treatment of SOE for 7 h at $18^{\circ}C$ was significantly increased but the intensity of H3K9me3 staining in PFFs was significantly decreased as compared with the control (p<0.05). Additionally, the level of histone acetylation in SCNT embryos at the zygote stage was significantly increased when reconstructed using SOE-treated cells (p<0.05), similar to that of IVF embryos at the zygote stage. The number of apoptotic cells was significantly decreased and pluripotency markers (Nanog, Oct4 and Sox2) were highly expressed in the blastocyst stage of SCNT embryos reconstructed using SOE-treated cells as nuclear donor (p<0.05). And there was observed a better development to the blastocyst stage in the SOE-treated group (p<0.05). Our results suggested that pre-treatment of cells with SOE could improve epigenetic reprogramming and the quality of porcine SCNT embryos.