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      • KCI등재

        개 REM134 유선종양세포의 sphere 형성을 통한 암 줄기세포 특성 분석

        변정수,이지현,정다운,구나연,조인수,차상호,Byeon, Jeong Su,Lee, Jienny,Jeong, Da-Un,Gu, Na-Yeon,Cho, In-Soo,Cha, Sang-Ho 대한수의학회 2018 大韓獸醫學會誌 Vol.58 No.4

        Canine mammary tumors are among the most frequently observed cutaneous tumors in female dogs. Cancer stem cells (CSCs), referred to as tumor-initiating cells, are thought to have properties similar to normal stem cells such as the ability to self-renewal and to differentiate into various cell types. Biological understanding of CSCs and the critical pathways involved in their maintenance are important in research and therapy for mammary tumors. We conducted the present study on sphere formation from REM134 cells by using methylcellulose to produce tumorspheres on a large scale and compared the specific markers of the spheres-formed and plating-cultured REM134 cells. The results revealed that the tumorspheres cultured in methylcellulose had higher seeding density and improved morphology compared to those produced in normal sphere formation medium. Expression levels of stemness markers and CSC-related markers were higher in tumorsphere-forming cells than in plating-cultured cells. Subsequently, we transplanted the tumorsphere-forming and plating-cultured cells into female nude mice to examine their tumorigenic potential. Tumor volume increased rapidly in mice transplanted with tumorsphere-derived cells compared to plating-cultured cells. We observed a novel sphere-forming condition for REM134 cells and showed that REM134 cell tumorspheres can exhibit improved CSC properties.

      • KCI등재

        Isolation and characterization of deer-derived mesenchymal stem cells

        Jeong Su Byeon(변정수),Na-Yeon Gu(구나연),Da-Un Jeong(정다운),Chan-Lan Kim(김찬란),In-Soo Cho(조인수),Jae-Young Song(송재영),Bang-Hun Hyun(현방훈),Jienny Lee(이지현) 한국예방수의학회 2019 예방수의학회지 Vol.43 No.3

        Mesenchymal stem cells (MSCs) are capable of differentiating into mesenchymal tissue such as bone, cartilage, muscle, and adipose, and have been isolated and characterized from various species. Deer adipose tissue-derived MSCs (dAD-MSCs) have not been studied and deer bone marrow-derived MSCs (dBM-MSCs) have not been fully characterized. In this study, we firstly isolated MSCs from deer tissues and then compared characteristics of dAD-MSCs and dBM-MSCs. dAD-MSCs and dBM-MSCs exhibited significant increase in proliferation under low-glucose DMEM culture condition during 20 and 10 passages consecutive passages, respectively. Both cells expressed cell surface markers such as CD73, CD90, and CD105, but did not express CD34 and CD45. Two types of cells expressed stemness markers (Oct4, Sox2, and Nanog) and exhibited differentiation potential into mesodermal lineages. Both cells exhibited osteogenic and chondrogenic differentiation potential, but poor adipogenic differentiation potential. Specifically, dAD-MSCs have a greater capacity for chondrogenic differentiation potential compared to dBM-MSCs. Collectively, we successfully isolated dAD-MSCs from deer for the first time. This study suggests that adipose tissue of deer could be used as a source of deer MSCs.

      • KCI등재

        개 유선종양세포에 대한 자연살해세포 독성

        정다운,변정수,구나연,정문희,김은희,김형석,조인수,송재영,현방훈,이지현,Jeong, Da-Un,Byeon, Jeong Su,Gu, Na-Yeon,Jung, Moonhee,Kim, Eun Hee,Kim, Hyung-Seok,Cho, In-Soo,Song, Jae-Young,Hyun, Bang-Hun,Lee, Jienny 대한수의학회 2020 大韓獸醫學會誌 Vol.60 No.1

        Natural killer (NK) cells play have a crucial role in the early phase of immune responses against various pathogens. We compared characteristics of canine NK cells against two canine mammary carcinoma cell lines, REM134 and CF41.Mg. REM134 showed higher expression of progesterone receptor, proliferative cell nuclear antigen, Ki67, multiple drug resistance, Bmi-1, c-myc, E-cadherin, and human epidermal growth factor receptor type-2 than that of CF41.Mg. For specific expansion and activation of NK cells, we isolated CD5 negative cells from canine peripheral blood mononuclear cells and co-cultured K562 cells in the presence of interleukin (IL)-2, IL-15, and IL-21 for 21 days. As a result, we found that expression markers of activated NK cells such as NKp30, NKp44, NKp46, NKG2D, CD244, perforin, granzyme B, and tumor necrosis factor alpha were highly upregulated. In addition, we found there was upregulated production of interferon gamma of activated NK cells against target cells such as REM134 and CF41.Mg. Specifically, we observed that cytotoxicity of NK cells against target cells was more sensitively reacted to CF41.Mg than REM134. Based on the results of this study, we recommend the development of an experimental application of CF41Mg, which has not been reported in canine mammary carcinoma research.

      • SCOPUSKCI등재

        $N_2O$ 분위기에서 열산화법으로 성장시킨 $SiO_2$초박막의 전기적 특성

        강석봉,김선우,변정수,김형준,Gang, Seok-Bong,Kim, Seon-U,Byeon, Jeong-Su,Kim, Hyeong-Jun 한국재료학회 1994 한국재료학회지 Vol.4 No.1

        $SiO_{2}$초박막(ultrathin film)의 두께 조절 용이성, 두께 균일성, 공정 재현성 및 전기적 특성을 향상시키기 위해 실리콘을 $N_{2}O$분위기에서 열산화시켰다. $N_2O$분위기에서 박막 성장시 산화와 동시에 질화가 이루어지기 때문에 전기적 특성의 향상을 가져올 수 있었다. 질화 현상에 의해 형성된 Si-N결합 형성은 습식 식각율과 ESCA분석으로 확인할 수 있었다. $N_2O$분위기에서 성장된 $SiO_{2}$박막은 Fowler-Nordheim(FN)전도 기구를 보여주었으며, 절열파괴 특성과 누설 전류특성 및 산화막의 신뢰성은 건식 산화막에 비해서 우수하였다. 또한 계면 포획밀도는 건식 산화막에 비해 감소하였고, 전하를 주입했을 때 생성되는 계면 준위의 양 또는 크게 감소하였다. 산화막 내부에서의 전하 포획의 양도 감소하였고, 전하를 주입하였을 때 생성되는 전하 포획의 양도 감소하였다. 이와 같은 전기적인 특성의 향상은 산화막 내부에서 약하게 결합하고 있는 Si-O 결합들이 Si-N결합으로의 치환과 스트레스 이완에 의하여 감소하였기 때문이다. The ultrathin oxide films less than 100$\AA$ were grown by thermal oxidation in $N_2O$ ambient to improve the controllability of thickness, thickness uniformity, process reproducibility and their electrical properties. Oxidation rate was reduced significantly at very thin region due to the formation of oxynitride layer in $N_2O$ ambient and moreover nitridation of the oxide layer was simultaneously accompanied during growth. The nitrogen incorporation in the grown oxide layer was characterized with the wet chemical etch-rate and ESCA analysis of the grown oxide layer. All the oxides thin films grown in $N_2O$, pure and dilute $O_2$ ambients show Fowler-Nordheim electrical conduction. The electrical characteristics of thin oxide films grown in $N_2O$ such as leakage current, electrical breakdown, interface trap density generation due to the injected electron and reliability were better than those in pure or dilute ambient. These improved properties can be explained by the fact that the weak Si-0 bond is reduced by stress relaxation during oxidation and replacement by strong Si-N bond, and thus the trap sites are reduced.

      • KCI등재

        하이드로젤 지지체 기반 3차원 환경에서 개 간엽줄기세포의 분화능 분석

        구나연,박미정,이지현,변정수,정다운,조인수,차상호,Gu, Na-Yeon,Park, Mi Jeong,Lee, Jienny,Byeon, Jeong Su,Jeong, Da-Un,Cho, In-Soo,Cha, Sang-Ho 대한수의학회 2018 大韓獸醫學會誌 Vol.58 No.4

        Mesenchymal stem cells (MSCs) are useful candidates for tissue engineering and cell therapy. Physiological cell environment not only connects cells to each other, but also connects cells to the extracellular matrix that provide mechanical support, thus exposing the entire cell surface and activating signaling pathways. Hydrogel is a polymeric material that swells in water and maintains a distinct 3-dimensional (3D) network structure by cross linking. In this study, we investigated the optimized cellular function for canine adipose tissue-derived MSCs (cAD-MSCs) using hydrogel. We observed that the expression levels of Ki67 and proliferating cell nuclear antigen, which are involved in cell proliferation and stemness, were increased in transwell-hydrogel (3D-TN) compared to the transwell-normal (TN). Also, transforming growth factor-${\beta}1$ and SOX9, which are typical bone morphogenesis-inducing factors, were increased in 3D-TN compared to the TN. Collagen type II alpha 1, which is a chondrocyte-specific marker, was increased in 3D-TN compared to the TN. Osteocalcin, which is a osteocyte-specific marker, was increased in 3D-TN compared to the TN. Collectively, preconditioning cAD-MSCs via 3D culture systems can enhance inherent secretory properties that may improve the potency and efficacy of MSCs-based therapies for bone regeneration process.

      • KCI등재

        Characteristics of mesenchymal stem cells derived from different tissues of goat

        Na-Yeon Gu(구나연),Da-Un Jeong(정다운),Jeong Su Byeon(변정수),Jae-Young Song(송재영),In-Soo Cho(조인수),Bang-Hun Hyun(현방훈),Jienny Lee(이지현) 한국예방수의학회 2019 예방수의학회지 Vol.43 No.3

        Mesenchymal stem cells (MSCs) are multipotent cells capable of replicating as undifferentiated cells, and have the potential of differentiating into mesodermal lineages. Goats are commonly used as animal models for bone tissue engineering to test the potential of stem cells for bone regeneration. Goat MSCs isolated from bone marrow (BM) or adipose tissue (AD) should be evaluated using in vitro assays, prior to their application in a tissue engineering project. In this study, we compared the stem cell properties of MSCs derived from goat AD, BM and ear skin tissue (ESK). As results, BM and ESK-MSCs exhibited a spindle-shaped morphology comparable to that of AD-MSCs. Especially, BM-MSCs could be cultured for significantly longer periods and exhibited the greatest expansion capacity, whereas AD-MSCs had the shortest culture time and lowest growth rate. Also, we compared differentiation potentials of AD, BM and ESK-MSCs into adipogenic, chondrogenic, and osteogenic lineages through specific staining and quantitative real-time RT-PCR. Collectively, we successfully isolated ESK-MSCs from goat for the first time. This study suggests that adult skin tissue of goat could be used as a source of goat MSCs. Further studies are needed to show the more information for establishment and fully characterization of goat ESK-MSCs.

      • KCI등재

        Lipopolysaccharide로 자극한 RAW 264.7 세포에서 성체줄기세포 유래 엑소좀(exosome)의 면역 조절 효과

        정수경 ( Soo Kyung Jung ),박미정 ( Mi Jeong Park ),이지현 ( Jienny Lee ),변정수 ( Jeong Su Byeon ),구나연 ( Na Yeon Gu ),조인수 ( In Soo Cho ),차상호 ( Sang Ho Cha ) 한국미생물생명공학회(구 한국산업미생물학회) 2016 한국미생물·생명공학회지 Vol.44 No.3

        본 연구는 대식세포에서 LPS를 이용하여 염증 유사 세포모델을 만들고 염증 유사 대식세포 모델에서 성체줄기세포의 면역 조절 능력을 평가하였다. LPS 자극에 의해 증가된IL-1β, TNF-α 및 IL-10의 생성은 성체줄기세포를 공배양한 실험군 뿐만 아니라 성체줄기세포를 배양한 상층 배양액을 처리한 실험군에서도 동일한 효과를 나타내었으며, 또한 성체줄기세포 유래 엑소좀을 염증 유사 대식세포 모델에 처리하여 유사한 결과를 관찰하였다. 이 결과는 성체줄기세포 자체의 염증 억제 기능보다는 성체줄기세포 유래 엑소좀을 포함하여 성체줄기세포가 분비하는 bioactive molecules에 의해 세포 간 신호 전달이 이루어지고 있음을 의미하며, 이러한 엑소좀은 염증 관련 질환 분야에 치료적 적용이 가능하고 또한 새로운 염증 치료제 개발의 툴로 사용될 수 있음을 시사한다. Mesenchymal stem cells (MSCs) are multipotent stem cells that can be differentiated into a variety of cell types, including adipocytes, osteoblasts, chondrocytes, β-pancreatic islet cells, and neuronal cells. MSCs have been reported to exhibit immunomodulatory effects in many diseases. Many studies have reported that MSCs have distinct roles in modulating inflammatory and immune responses by releasing bioactive molecules. Exosomes are cell-derived vesicles present in biological fluids, including the blood, urine, and cultured medium of cell cultures. In this study, we investigated the immunomodulatory effects of mouse adipose tissue-derived MSCs (mAD-MSCs), cultured medium (MSC-CM) of mAD-MSCs, and mAD-MSCderived exosomes (MSC-Exo) on lipopolysaccharide (LPS)-induced RAW 264.7 cells. We observed that the expression levels of IL-1β, TNF-α, and IL-10 were significantly increased in LPS-stimulated RAW 264.7 cells compared to those in LPS-unstimulated RAW 264.7 cells. Additionally, these values were significantly (p < 0.05) decreased in mAD-MSCs-RAW 264.7 cell co-culture groups, MSC-CM-treated groups, and MSC-Exotreated groups. MSCs can modulate the immune system in part by secreting cytokines and growth factors. We observed that immunomodulatory factors such as IL-1β, TNF-α, and IL-10 were secreted by mAD-MSCs under co-culturing conditions of mAD-MSCs with activated RAW 264.7 cells. In addition, mAD MSC-derived exosomes exhibited similar immunomodulatory effects in activated RAW 264.7 cells. Therefore, our results suggest that mAD-MSCs have an immunomodulatory function through indirect contact.

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