• BMB Reports
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• v.52 no.6
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• pp.403-408
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• 2019

Dorsoventral patterning of body axis in vertebrate embryo is tightly controlled by a complex regulatory network of transcription factors. Ventx1.1 is known as a transcriptional repressor to inhibit dorsal mesoderm formation and neural differentiation in Xenopus. In an attempt to identify, using chromatin immunoprecipitation (ChIP)-Seq, genome-wide binding pattern of Ventx1.1 in Xenopus gastrulae, we observed that Ventx1.1 associates with its own 5'-flanking sequence. In this study, we present evidence that Ventx1.1 binds a cis-acting Ventx1.1 response element (VRE) in its own promoter, leading to repression of its own transcription. Site-directed mutagenesis of the VRE in the Ventx1.1 promoter significantly abrogated this inhibitory autoregulation of Ventx1.1 transcription. Notably, Ventx1.1 and Xcad2, an activator of Ventx1.1 transcription, competitively co-occupied the VRE in the Ventx1.1 promoter. In support of this, mutation of the VRE down-regulated basal and Xcad2-induced levels of Ventx1.1 promoter activity. In addition, overexpression of Ventx1.1 prevented Xcad2 from binding to the Ventx1.1 promoter, and vice versa. Taken together, these results suggest that Ventx1.1 negatively regulates its own transcription in competition with Xcad2, thereby fine-tuning its own expression levels during dorsoventral patterning of Xenopus early embryo.

• Bulletin of the Korean Chemical Society
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• v.34 no.4
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• pp.1175-1180
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• 2013

A microfabricated electrochemical cell comprising PDMS-based microchannel and in-channel gold microelectrodes was fabricated as a sensitive and a miniature alternative to the conventional electroanalytical systems. A reproducible fabrication procedure enabled patterning of multiple microelectrodes integrated within a PDMS-based fluidic network. The active area of each electrode was $200{\mu}m{\times}200{\mu}m$ with a gap of $200{\mu}m$ between the electrodes which resulted in a higher signal to noise ratio. Also, the PDMS layer served the purpose of shielding the electrical interferences to the measurements. Analytes such as potassium ferrocyanide; amino acid: cysteine and nucleoside: guanosine were characterized using the fabricated cell. The microchip was comparable to bulk electrochemical systems and its applicability was also demonstrated with flow injection based rapid amperometric detection of DNA samples. The device so developed shall find use as a disposable electrochemical cell for rapid and sensitive analysis of electroactive species in various industrial and research applications.

• Journal of Microbiology and Biotechnology
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• v.25 no.3
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• pp.381-385
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• 2015

In the present study, drop-on-demand two-dimensional patterning of unstained and stained bacterial cells on untreated clean wafers was newly conducted using an electrospray pulsed jet. We produced various spotted patterns of the cells on a silicon wafer by varying the experimental conditions, such as the frequency, flow rate, and translational speed of the electrospray system in a two-dimensional manner. Specifically, the electrospray's pulsed jet of cell solutions produced alphabetical patterns consisting of spots with a diameter of approximately $10{\mu}m$, each of which contained a single or a small number of viable bacteria. We tested the viability of the patterned cells using two visualization methods. This pattering technique is newly tested here and it has the potential to be applied in a variety of cell biology experiments.

• Journal of Biomedical Engineering Research
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• v.40 no.5
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• pp.165-170
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• 2019

Cancer cells are different from normal cells in terms of life cycle, behavior, and growth patterns. Cancer cells can migrate freely in the body through blood vessels and lymph nodes. The cancer cells easily interact with various substrates including extracellular matrix and vessels and they can differentiate in the new environment. However, it is not well known about the adhesion preference of cancer cells on the substrate and the mechanism of their interaction. In this study, we prepared the nano-, micro-patterned substrates using E-beam lithography techniques. MCF-7 cells were tested on the substrates to find out their adhesion preference. The substrates were made by polydimethylsiloxane (PDMS) with specific patterns including pillars with a diameter of 500 nm, 700 nm, $3{\mu}m$ and $5{\mu}m$. MCF-7 cells were seeded on the substrates and incubated for 24 hours. As a result, this study clearly demonstrated that the MCF-7 cells preferred 700 nm patterning.

• Molecules and Cells
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• v.45 no.4
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• pp.243-256
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• 2022

Transcriptional regulation, a core component of gene regulatory networks, plays a key role in controlling individual organism's growth and development. To understand how plants modulate cellular processes for growth and development, the identification and characterization of gene regulatory networks are of importance. The SHORT-ROOT (SHR) transcription factor is known for its role in cell divisions in Arabidopsis (Arabidopsis thaliana). However, whether SHR is involved in hypocotyl cell elongation remains unknown. Here, we reveal that SHR controls hypocotyl cell elongation via the transcriptional regulation of XTH18, XTH22, and XTH24, which encode cell wall remodeling enzymes called xyloglucan endotransglucosylase/hydrolases (XTHs). Interestingly, SHR activates transcription of the XTH genes, independently of its partner SCARECROW (SCR), which is different from the known mode of action. In addition, overexpression of the XTH genes can promote cell elongation in the etiolated hypocotyl. Moreover, confinement of SHR protein in the stele still induces cell elongation, despite the aberrant organization in the hypocotyl ground tissue. Therefore, it is likely that SHR-mediated growth is uncoupled from SHR-mediated radial patterning in the etiolated hypocotyl. Our findings also suggest that intertissue communication between stele and endodermis plays a role in coordinating hypocotyl cell elongation of the Arabidopsis seedling. Taken together, our study identifies SHR as a new crucial regulator that is necessary for cell elongation in the etiolated hypocotyl.

• KSBB Journal
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• v.23 no.3
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• pp.263-270
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• 2008

A poly-(dimethylsiloxane) (PDMS) surface modified by the successive deposition of the polyelectrolytes, poly-(allylamine hydrochloride) (PAH), poly-(diallyldimethylammoniumchloride) (PDAC), poly-(4-ammonium styrenesulfonic acid) (PSS), and poly-(acrylic acid) (PAA), was presented for the application of selective cell immobilization. It is formed via electrostatic attraction between adjacent layers of opposite charge. The modified PDMS surface was examined using static contact angle measurements and fourier transform infrared (FT-IR) spectrophotometer. The wettability of the PDMS surface could be easily controlled and functionalized to be biocompatible through regulation of layer numbers. The modified PDMS surface provides appropriate environment for adhesion to cells, which is essential technology for cell patterning with high yield and viability in the patterning process. This method is reproducible, convenient, and rapid. It could be applied to the fabrication of biological sensing, patterning, microelectronics devices, screening system, and study of cell-surface interaction.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.118-118
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• 2002

Programmed cell death (PCD) is thought as a well-controlled process by which unwanted cells are selectively eliminated. During the last decade many researches have elucidated molecules and their interactions involved in cell death by using largely in vitro induction of cell death or survival signals in a more defined manner, While these critical information and novel findings provide us with clearer understanding of mechanisms underlying cell death, it does by no means explain how PCD occurs and which cells or tissues are affected during normal embryonic development in vivo. In this study, we used zebrafish to examine whether the PCD is occurring selectively or randomly in developing embryos by whole mount in situ TUNEL analysis with specific markers for neural cells. The result revealed that the degree and distribution of TUNEL staining varied considerably throughout gastrulation stage, and there was also a number of TUNEL-negative embryos. Most of TUNEL-positive cells were scattered randomly throughout the blastoderm. During the gastrulation stage about 75 % of the embryos analyzed exhibited more than 5 TUNEL-positive cells. As the dorsal epiblast begins to thicken rather abruptly near the end of gastrulation, TUNEL-positive cells were mainly located along the dorsal side. Although there were some variations in TUNEL staining during segmentation and pharyngeal stages, TUNEL staining continued to be localized to the central nervous system, and was also detected in the sensory organs, trigeminal ganglions, and the primary sensory neurons. High levels of the cell death in developing brain between 20-somite and prim-6 stages are thought to play a role in the morphogenesis and organization of the brain. At prim-16 stage, cell death is considerably reduced in the brain region. Dying cells are mainly localized to the prospective brain region where ectodermal cells are about to initiate neurogenesis. As development progressed, high levels and more reproducible patterns of cell death were observed in the developing nervous system. Intensive TUNEL staining was restricted to the trigeminal ganglions, the primary sensory neurons, and sensory organs, such as olfactory pits and otic vesicles. Thus, PCD patterning in zebrafish embryos occurs randomly at early stages and becomes restricted to certain region of the embryos. The spatio-temporal pattern of PCD during the early embryonic development in zebrafish will provide basic information for further studies to elucidate genes involved in. regulation of PCD largely unknown in vivo during vertebrate embryogenesis.

• Molecules and Cells
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• v.39 no.4
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• pp.352-357
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• 2016

Vertebrate neurogenesis requires inhibition of endogenous bone morphogenetic protein (BMP) signals in the ectoderm. Blocking of BMPs in animal cap explants causes the formation of anterior neural tissues as a default fate. To identify genes involved in the anterior neural specification, we analyzed gene expression profiles using a Xenopus Affymetrix Gene Chip after BMP-4 inhibition in animal cap explants. We found that the xCyp26c gene, encoding a retinoic acid (RA) degradation enzyme, was upregulated following inhibition of BMP signaling in early neuroectodermal cells. Whole-mount in situ hybridization analysis showed that xCyp26c expression started in the anterior region during the early neurula stage. Overexpression of xCyp26c weakly induced neural genes in animal cap explants. xCyp26c abolished the expression of all trans-/cis-RA-induced posterior genes, but not basic FGF-induced posterior genes. Depletion of xCyp26c by morpholino-oligonucleotides suppressed the normal formation of the axis and head, indicating that xCyp26c plays a critical role in the specification of anterior neural tissue in whole embryos. In animal cap explants, however, xCyp26c morpholinos did not alter anterior-to-posterior neural tissue formation. Together, these results suggest that xCyp26c plays a specific role in anterior-posterior (A-P) neural patterning of Xenopus embryos.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.37-42
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• 2002

In ascidians, a primitive chordate, maternal cytoplasmic factors and inductive interactions are involved in the specification of cell fate in early embryos. The larval structure of ascidians is relatively simple, and the major mesodermal tissues of the tadpole larva are notochord, muscle and mesemchyme. Formation of muscle cells is a cell-autonomous process, and localized maternal macho-1 mRNA specify muscle fate in the posterior marginal zone of the early embryo. In contrast, inductive influence from endoderm precursors plays important roles in the specification of notochord and mesenchyme fates. FGF-Ras-MAPK signaling is involved in the induction of both tissues. The difference in responsiveness of the posterior mesenchyme and anterior notochord precursors is caused by the presence or absence of the posterior-vegetal egg cytoplasm, respectively. In these cases, directed signal may polarizes the responding cells and cause asymmetric cell divisions that operate in both the anterior and posterior regions.

• Journal of Biomedical Engineering Research
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• v.31 no.2
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• pp.87-93
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• 2010

Researches for stem cells have been focused on scientists in biomedical sciences as well as clinical application for its great therapeutic potentials. Stem cells have two distinct characteristics: self-renewal and differentiation. In this short review, the links between stem cell research and biomedical engineering is discussed based on the basic characteristics of stem cells. This concept can be extended to the fundamental questions of biological sciences for cells such as proliferation, apoptosis, differentiation, and migration. For understanding proliferation and apoptosis of stem cells, techniques from biomedical engineering such as surface patterning, MEMS, nanotechnologies have been used. The advanced technologies such as microfluidic technologies, three dimensional scaffold fabrication, and mechanical/electrical stimulation have also been used in cell differentiation and migration. Basic and unsolved questions in the stem cell research field have limitations by studying conventional technologies. Therefore, the strategic fusion between stem cell biology and novel biomedical engineering field will break the barriers for understanding fundamental questions of stem cells, which can open the window for the clinical applications of stem cell based therapeutics as well as regeneration of damaged tissues.