Bacillus subtilis CH-10에 의한 폴리감마글루탐산의 생산에 관한 연구

구나연,김춘희,김병우,남수완,권현주,김동은,김영만,전숭종,Gu Na-Yeon,Kim Choon-Hee,Kim Byung-Woo,Nam Soo-Wan,Kwon Hyun-Ju,Kim Dong-Eun,Kim Young-Man,Jeon Sung-Jong 한국생명과학회 2006 생명과학회지 Vol.16 No.2

A bacterium that produced a large amount of poly-$\gamma$-glutamate (PGA) was isolated from the compost and designated as Bacillus subtilis CH-10. The optimum temperature and pH for PGA production were at $37^{\circ}C$ and 7.5, respectively. The maximum amount of PGA production (18.84 mg/ml) was obtained when it was grown in a medium containing 3% L-glutamate and 5% sucrose at $37^{\circ}C$ with shaking. The result that the L-glutamate significantly induced PGA production indicates that it produces a PGA by the glutamate dependent manner. Some properties of the PGA obtained at different times of cultivation were investigated by SDS-PAGE and ninhydrin analysis. The PGA production was elongated along with cultivation time and maximum amount was achieved at 96 h. Average molecular weight of PGA was estimated to be 1100 kDa by FDNB method. 폴리감마글루탐산(PGA)을 대량생산하는 균주를 축분퇴비에서 분리하고 생리 생화학적 특징 및 16s rRNA 염기서열을 분석한 결과, Bacillus subtilis로 동정하고 strain CH-10 으로 명명하였다. PGA 생산을 위한 배지의 최적 온도와 pH는 각각 $37^{\circ}C$, 7.5 이었다. 또한 배지성분 중에서 PGA 생산에 최적인 탄소원과 질소원은 각각 5% sucrose와 3% L-glutamate인 것으로 나타났다. 이 균주는 L-glutamate에 의해 PGA 생산이 대량으로 유도되었고 따라서 질소원으로 L-glutamate를 반드시 필요로 하는 glutamate 의존성 균주인 것으로 사료되었다. 배양시간에 따라 생산된 PGA의 생화학적 특정을 SDS-PAGE와 ninhydrin 분석을 통하여 검토하였다. PGA의 생산량은 배양시간에 따라 증가하였고 배양 후 96시간에서 최고 생산량을 나타내었다, 생산된 PGA의 평균분자량을 FDNB법으로 분석한 결과, 1,100 kDa인 것으로 나타났다.

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.

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.

구타이미술(具?美術)과 일본 전후(戰後) 모더니티의 경험

구나연(Gu Na Yeon) 한국미술사교육학회 2011 美術史學 Vol.- No.25

본 논문은 일본의 구타이미술협회(具?美術協會, 1954-1972 이하 ‘구타이’라 칭함)가 전후 모더니티를 경험하는 방식과 그것이 어떻게 이들의 작품에 반영되고 있는가에 관한 것이다. 본 논문은 크게 세 가지 문제를 다루고 있는데, 첫째 초기 구타이미술에 나타난 모더니티의 성격, 둘째 초기 구타이가 어떤 외부적 동인에 의해 회화그룹으로 전환되었는가, 셋째 구타이회화와 정치적 이데올로기와의 관계가 그것이다. 구타이가 경험한 전후 모더니티의 역사는 산업 생산성을 기반으로 한 자본주의와 미국 중심의 외교로 형성된 냉전하의 민주주의가 급속하게 진행되는 과정이었다. 따라서 이들의 미술에는 패전으로 상징되는 근대적 주체로부터의 이탈과 동시에 새로운 현대적 주체에 대한 인식의 교차 안에서 근원적인 변화에 대한 욕망과 체험이 담겨있고, 구타이의 활동에서 찾을 수 있는 미술사적 역류 역시 전후 일본이 제시하고 있는 보수와 혁신의 긴장, 미국의 세계전략 등의 현실적 영향들에서 자유로울 수 없었던 것을 보여준다. 더욱이 냉전체제 속에서 미국은 추상미술을 통해 미국적 자유이상의 가치를 문화적으로 선전하려는 열망을 가지고 있었고, 일본은 재벌 중심의 경제부흥과 자민당 출범과 같은 정치적 사건을 통해 보수적 민주주의라는 현대 사회의 근본 형태를 수립하게 된다. 따라서 구타이가 바로 이 시기 일본 앵포르멜의 대표자로서 활동했다는 것은 구타이가 설사 미처 인식하지 못했다 하더라도 구타이의 회화가 일본 사회가 수립한 보수적 민주주의의 가치와 냉전체제의 이데올로기를 적극적으로 수용했다는 정치성을 드러낸다. This paper is about the Gutai(1954-1972) and post-war Japanese modernity. It deals with three questions. Firstly, it is how modernity related in the Gutai and their works. Secondly, why the Gutai had to change to painting group. And lastly, what is the Gutai in modern Japanese political ideology in post-war. The modern history of post-war Japan in the Gutai had some connection with capitalism, democracy and cold war. Thur the Gutai art is the symbol that the modern subject(in pre-war) move toward the comtemporary subject in Japan. It is the desire for radical changes to advanced modernity. So flowing backward of Gutai on art history could explain the tension of the political conservatism between innovation and Socialism between American liberal propaganda in cold war. Besides it regarded the Informel painting and the abstract expressionism in same light as democracy. It rises to a political issues and high grows of Japanese economy. It is remarkable that the Gutai in this period was the best well-known art group of Japan. Because the Gutai embraced the conservative democracy and the ideology of cold war. And it is the reason of the changes from a radical experimental art group to a common painting group for the Gutai.

전기 자극에 의한 개 연골세포의 특성 강화

구나연(Na-Yeon Gu),이세아(Se-A Lee),정소연(So Yeon Jeong),이지현(Jienny Lee),이윤희(Yoon-Hee Lee),현방훈(Bang-Hun Hyun) 한국생물공학회 2020 KSBB Journal Vol.35 No.3

Electrical stimulation (ES) is known to guide the development and regeneration of many tissues. Cartilage has no direct blood supply, thus it has little or no capacity to repair itself. Articular cartilage is to provide a smooth lubricating surface to the joints and facilitates the transfer of loads with a low frictional coefficient. Meniscus is cartilaginous tissues that provide structural integrity to the knee when it undergoes tension and torsion. We tried to isolate and characterize chondrocytes from articular cartilage (Ar-chon) and meniscus (Me-chon) of dogs. Also, we exposed Me-chon to ES and assessed changes in the chondrogenic potential. As results, Ar-chon and Me-chon had fibroblasts-like morphology. The proliferative capacity of Me-chon was higher than those of Ar-chon in the early passage. Both cells expressed cell surface markers such as CD29, CD44, and CD90, but did not express CD34, CD117, and MHC-2. ES was applied to Mechon cultured under condition of high-density micromass under ES of 10 V/㎝, with duration of 10 ㎳ and a frequency of 2 ㎐ for three days. And then, gene expression of chondrogenic markers such as type II collage alpha 1, Aggrecan, and SOX9 was analyzed. As a result, we observed the upregulated expression level of chondrogenic markers of Me-chon by specific ES in absence of exogenous factors. Also, we found that ES significantly upregulated expression level of heat shock protein, which affects cartilage formation. Taken together, ES-driven Me-chon is thought to be useful for basic and therapeutic research applications in the early passage.

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.

Effects of canine mesenchymal stem cells-derived exosomes in a mouse model of canine mammary tumor

정소연(So Yeon Jeong),이세아(Se-A Lee),구나연(Na-Yeon Gu),이지현(Jienny Lee),이윤희(Yoon-Hee Lee),현방훈(Bang-Hun Hyun) 한국예방수의학회 2020 예방수의학회지 Vol.44 No.4

Canine mammary tumors account for ~30% of all tumors in the female dogs and approximately 50% of the tumors are malignant. Exosomes have been the focus of great interest, as they appear to be involved in numerous important cellular processes. In this study, we examined the anti-tumor effects of canine mesenchymal stem cells-derived exosomes (MSC-exosomes) in an experimental murine mammary tumor model using canine mammary carcinoma cells, REM134. The MSC-exosomes were injected tumor site and tail vein of REM134 xenografted mice. We found that tumor size of the MSC-exosomes-treated group decreased compared to those of the only tumor group in REM134-driven tumorigenic mouse model. In addition, the MSC-exosomes-treated tumor group showed meaningfully reduced expression levels of the MMP-3, IL-1β, IL-6, and TNF-α compared to those in the tumor group. Specifically, we confirmed that the expression level of the CD133, potent cancer stem cell (CSC) markers, decreased in the MSC-exosomes-treated tumor group compared to the tumor group. This study suggests that the MSC-exosomes exhibited anti-tumor effects through downregulating CSC-related markers in the canine mammary tumor murine model. Further study is needed in the future, and we are conducting research on the detailed anti-tumor mechanism of the MSC-exosomes.

전기 자극을 이용한 CD271 양성 중간엽줄기세포의 연골세포 분화능

정소연(So Yeon Jeong),이세아(Se-A Lee),구나연(Na-Yeon Gu),임성인(Seong-In Lim),이지현(Jienny Lee),이윤희(Yoon-Hee Lee),현방훈(Bang-Hun Hyun) 한국생물공학회 2021 KSBB Journal Vol.36 No.1

Mesenchymal stem cells (MSCs) exhibit self-renewing capacity and multi-lineage differentiation potential. CD271 has been identified as a marker of the most homogeneous MSCs subset. In this study, we separated CD271 positive MSCs (CD271-MSCs) from canine adipose tissue-derived MSCs (AD-MSCs) using immunomagnetic selection method. Both CD271-MSCs and AD-MSCs showed similar morphology and expressed common immunophenotypic markers (CD34, CD44, and CD45). However, CD271-MSCs and AD-MSCs exhibited difference in chondrogenic differentiation potential. Specifically, CD271-MSCs exhibited increased expression levels of chondrocyte specific markers, collagen type II alpha 1 (COL2A1) and Aggrecan compared to AD-MSCs. Also, electrical stimulation (ES)-treated CD271-MSCs have a greater capacity for expressing heat shock protein 70 (HSP70), COL2A1, and Aggrecan compared to control without exogenous differentiation condition. This study might contribute to the differentiation potential of homogenous MSCs into cartilage cells by specific ES condition in the absence of exogenous differentiation agents.

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.

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.