• The Journal of the Korea Contents Association
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• v.13 no.8
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• pp.109-117
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• 2013

This research is for selection of animal and establishment of guideline which for animal robot design for the purpose of emotionally disturbed children's psychotherapy. In the case of emotionally disturbed children because of poor expression about themselves's emotional condition, there are many restriction in the existing cure and educational method. Using research method of animal shape robot which is starting to make attempt since the late 1990s is effective to learning disturbed children. For the development of animal robot for psychotherapy of emotionally disturbed children first of all which animal is selected is very important. Therefore at the early stage of research preference surveyed to children including emotionally disturbed children and decide an object of animal robot. And what kind of shape is very preferred is also preference surveyed. Through focusing group interview about shape, color, facial expression, sound, and move which have an influence on outward shape design of robot, present the guideline which is needed to animal robot design for emotionally disturbed children.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.216-216
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• 2004

Genetically modified domestic animals have many potential applications ranging from basic research to production agriculture. One of the goals in transgenic animal production schemes is to reliably predict the expression pattern of the foreign gene. Establishing a method to screen genetically modified embryos for transgene expression before transfer to surrogates may improve the likelihood of producing offspring with the desired expressing pattern. (omitted)

• Animal cells and systems
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• v.3 no.2
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• pp.181-185
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• 1999

Xenopus oocyte is one of the widely used heterologous expression systems of ion channels for electrophysiological studies. Here we describe a new method in which cRNA produced by polymerase chain reaction (PCR) and in vitro transcription is injected to express ion channels in oocytes. This method enables us (1) to eliminate all or a part of the untranslated region of the cDNA and to replace it with a known sequence which helps increase the expression level in oocytes, and (2) to use the PCR product for in vitro transcription without subcloning. Using this method, the expression level of one of the neuronal nicotinic acetylcholine receptors (nAChRs) $\alpha$$_{6}$ subtype in oocytes was systematically increased by more than 100-fold, which was confirmed both by the $\alpha$-Bungarotoxin ($\alpha$,/TEX>Bgt) binding assay and the current measurement.t.

• Journal of Animal Reproduction and Biotechnology
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• v.39 no.1
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• pp.2-11
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• 2024

Background: The small intestine plays a crucial role in animals in maintaining homeostasis as well as a series of physiological events such as nutrient uptake and immune function to improve productivity. Research on intestinal organoids has recently garnered interest, aiming to study various functions of the intestinal epithelium as a potential alternative to an in vivo system. These technologies have created new possibilities and opportunities for substituting animals for testing with an in vitro model. Methods: Here, we report the establishment and characterisation of intestinal organoids derived from jejunum tissues of adult pigs. Intestinal crypts, including intestinal stem cells from the jejunum tissue of adult pigs (10 months old), were sequentially isolated and cultivated over several passages without losing their proliferation and differentiation using the scaffold-based and three-dimensional method, which indicated the recapitulating capacity. Results: Porcine jejunum-derived intestinal organoids showed the specific expression of several genes related to intestinal stem cells and the epithelium. Furthermore, they showed high permeability when exposed to FITC-dextran 4 kDa, representing a barrier function similar to that of in vivo tissues. Collectively, these results demonstrate the efficient cultivation and characteristics of porcine jejunum-derived intestinal organoids. Conclusions: In this study, using a 3D culture system, we successfully established porcine jejunum-derived intestinal organoids. They show potential for various applications, such as for nutrient absorption as an in vitro model of the intestinal epithelium fused with organ-on-a-chip technology to improve productivity in animal biotechnology in future studies.

• Proceedings of the Korean Society for Bioinformatics Conference
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• 2003.10a
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• pp.170-177
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• 2003

Gene expression data are the quantitative measurements of expression levels and ratios of numberous genes in different situations based on microarray image analysis results. The process to draw meaningful information related to genomic diseases and various biological activities from gene expression data is known as gene expression data analysis. In this paper, we present a hierarchical clustering method of gene expression data based on self organizing map which can analyze the clustering result of gene expression data more efficiently. Using our proposed method, we could eliminate the uncertainty of cluster boundary which is the inherited disadvantage of self organizing map and use the visualization function of hierarchical clustering. And, we could process massive data using fast processing speed of self organizing map and interpret the clustering result of self organizing map more efficiently and user-friendly. To verify the efficiency of our proposed algorithm, we performed tests with following 3 data sets, animal feature data set, yeast gene expression data and leukemia gene expression data set. The result demonstrated the feasibility and utility of the proposed clustering algorithm.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.2
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• pp.163-170
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• 2020

Partheno Embryo's research is known to play a very important role in identifying the development of embryonic cells or analyzing the genetic mechanisms of embryonic development, but the information on apoptosis formed during the early stage of development on Partheno Embryo is very little. Therefore, this study analyzed whether the embryonic cell death of unit embryos can be inhibited by adding Scriptaid, one of HDACi, which plays a role in demethylation of histone proteins as a method of regulating the cell cycle in the early embryo development of Partheno Embryo. As a result, the differentiation rate was higher in the group that added Scriptaid and FBS, but the cellular development was higher in the group that added pregnant serum to Scriptaid. As a result of analyzing the expression of the gene through IF and PCR, the group with the addition of gestational serum increased the expression of BCL2 and PCNA, which affects the anti-Casp3 action in cell survival. In addition, it is interpreted that treatment of Scriptaid for 16 hours, rather than 24 h treatment lowers the expression of Casp-3, a representative factor of apoptosis, and also increases embryonic development, thus affecting early embryo development. Therefore, it is concluded that the 16-hour treatment of Scriptaid and the use of gestational serum will inhibit cell death in the early embryonic development and increase the development rate of the embryo.

• Journal of Veterinary Clinics
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• v.36 no.6
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• pp.319-324
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• 2019

Receptor tyrosine kinases (RTK) are major promising targets in anticancer therapy in human and veterinary medicine. Using immunohistochemistry method, we evaluated the expressionof five types RTK (PDGFR-α, PDGFR-β, VEGFR 2, c-Kit and Abl) in the six canine brain tumor samples (2 meningioma, 2 astrocytoma, 1 ependymoma and 1 choroid plexus papilloma). A total of five samples expressed PDGFR-β (5/6), one sample, the choroid plexus papilloma, expressed c-Kit (1/6), and a total of two samples expressed Abl (2/6). None of the samples showed expression of PDGFR-α and VEGFR 2. We demonstrate that a significant portion of canine brain tumors express tyrosine receptors for growth factors and show that these receptors generally localize to tumor cell membranes and the cytoplasm. Evaluation of immunohistochemical expression for the RTKs PDGFR-β, c-Kit, and Alb in canine brain samples reveals an interesting potential for molecular targeting by TKIs in therapeutic studies of canine brain tumors, and more studies will be needed to assess the interactions and efficacy of these RTKs and TKIs. Based on these results, we have some evidence for novel chemotherapeutic trials using TKIs for canine nervous tumors.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.4
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• pp.489-498
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• 2018

Objective: Foot and mouth disease (FMD) and porcine reproductive and respiratory syndrome (PRRS) are major diseases that interrupt porcine production. Because they are viral diseases, vaccinations are of only limited effectiveness in preventing outbreaks. To establish an alternative multi-resistant strategy against FMD virus (FMDV) and PRRS virus (PRRSV), the present study introduced two genetic modification techniques to porcine cells. Methods: First, cluster of differentiation 163 (CD163), the PRRSV viral receptor, was edited with the clustered regularly interspaced short palindromic repeats-CRISPR-associated protein 9 technique. The CD163 gene sequences of edited cells and control cells differed. Second, short hairpin RNA (shRNAs) were integrated into the cells. The shRNAs, targeting the 3D gene of FMDV and the open reading frame 7 (ORF7) gene of PRRSV, were transferred into fibroblasts. We also developed an in vitro shRNA verification method with a target gene expression vector. Results: shRNA activity was confirmed in vitro with vectors that expressed the 3D and ORF7 genes in the cells. Cells containing shRNAs showed lower transcript levels than cells with only the expression vectors. The shRNAs were integrated into CD163-edited cells to combine the two techniques, and the viral genes were suppressed in these cells. Conclusion: We established a multi-resistant strategy against viral diseases and an in vitro shRNA verification method.

• Journal of Embryo Transfer
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• v.31 no.3
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• pp.169-177
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• 2016

There is a growing interest in the application of primary hepatocytes for treatment of liver diseases in humans and for drug development. Several studies have focused on long-term survival and di-differentiation blocking of primary hepatocytes in an in vitro culture system. Therefore, the present study also aimed to optimize an in vitro culture system using primary rat hepatocytes. Primary rat hepatocytes from 6-week-old male Crl:CD rats were isolated using a modified two-step collagenase perfusion. Healthy $3.5{\times}10^6$ primary rat hepatocytes were seeded into a 2 dimensional (2D) culture in a 25T culture flask coated with collagen type I or into a 3D culture in a 125-ml spinner flask for 7 days. Production of plasma protein (ALB and TF), apoptosis (BAX and BCL2), and CYP (CYP3A1) related genes were compared between the 2D and 3D culture systems. The 3D culture system had an advantage over the 2D system because of the relatively high expression of ALB and low expression of BAX in the 3D system. However, the level of CYP3A1 did not improve in the 3D culture with and without the presence of a dexamethasone inducer. Therefore, 3D culture has an advantage for albumin production and primary rat hepatocyte survivability, but a low expression of CYP3A1 indicated that primary rat hepatocytes require a high-density culture for stress reduction by continuous flow.

• Reproductive and Developmental Biology
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• v.36 no.1
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• pp.13-19
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• 2012

In order to provide the basis for developing practical mouse embryonic stem cells (mESCs) culture method, how the endogenous level of self-renewal-stimulating factor genes was altered in the mESCs by different extracellular signaling was investigated in this study. For different extracellular signaling, mESCs were cultured in 2 dimension (D), 3D and integrin-stimulating 3D culture system in the presence or absence of leukemia inhibitory factor (LIF) and transcriptional level of $Lif$, $Bmp4$ and $Wnt3a$ was evaluated in the mESCs cultured in each system. The expression of three genes was significantly increased in 3D system relative to 2D system under LIF-containing condition, while only $Wnt3a$ expression was increased by 3D culture under LIF-free condition. Stimulation of integrin signaling in mESCs within 3D system with exogenous LIF significantly up-regulated transcriptional level of $Bmp4$, but did not induce transcriptional regulation of $Lif$ and $Wnt3a$. In the absence of LIF inside 3D system, the expression of $Lif$ and $Bmp4$ was significantly increased by integrin signaling, while it significantly decreased $Wnt3a$ expression. Finally, the signal from exogenous LIF significantly caused increased expression of $Lif$ in 2D system, decreased expression of $Bmp4$ in both 2D and 3D system, and decreased expression of $Wnt3a$ in integrin-stimulating 3D system. From these results, we identified that endogenous expression level of self-renewal-stimulating factor genes in mESCs could be effectively regulated through artificial and proper manipulation of extracellular signaling. Moreover, synthetic 3D niche stimulating endogenous secretion of self-renewal-stimulating factors will be able to help develop growth factor-free maintenance system of mESCs.