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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.
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.
2016년에서 2018년에 국내 말 인플루엔자 백신 접종 후 항체 양성률
조민수,이주연,이상규,송재영,이지현,현방훈,조수동,우인옥,Cho, Min-Su,Lee, Ju-Yeon,Lee, Sang Kyu,Song, Jae Young,Lee, Jienny,Hyun, Bang-Hun,Cho, Soo-Dong,Ouh, In-Ohk 대한수의학회 2019 大韓獸醫學會誌 Vol.59 No.3
Equine influenza (EI) is the main cause of respiratory illness in equines across the globe and is caused by equine influenza A virus (EIV-A), which has impacted the equine industry internationally because of the marginal mortality and high morbidity. In the present study, the immune responses after equine influenza vaccination were evaluated in 4,144 horses in Korea using the hemagglutination inhibition (HI) assay. The equine influenza virus (EIV), A/equine/South Africa/4/03 (H3N8), was used as the antigen in the HI assay. The mean seropositive rates were 89.2% (97.4% in 2016, 77.6% in 2017, and 92.4% in 2018). This paper highlights the advances in understanding the effects of vaccines and control strategies for mitigating the emerging menace by EIV.
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.
Vegetable Peptones의 세포증식 및 콜라겐생성 촉진효과
정은선 ( Eun Sun Jung ),이종성 ( Jong Sung Lee ),이지현 ( Jienny Lee ),허성란 ( Sung Ran Huh ),김영수 ( Young Soo Kim ),황왕택 ( Wang Taek Hwang ),박덕훈 ( Deok Hoon Park ) 대한화장품학회 2009 대한화장품학회지 Vol.35 No.1
피부노화의 주요 원인으로는 타입 I 콜라겐 생합성 저하 및 피부 진피세포의 증식 활성 감소를 들 수 있다 효과적으로 피부노화를 관리하기 위해서는 안전하면서도 효능이 우수한 소재를 찾는 것이 필요하다 이를 위해 본 연구진은 항노화소재를 스크리닝하였고 완두콩과 밀 펩톤이 성체줄기세포의 세포증식을 증가시키는 효능을 관찰하였다 완두콩과 밀 펩톤을 포함하는 식물성 펩톤은 다양한 크기의 펩타이드와 아미노산을 함유하고 있어 세포배양 시 첨가해 주면 영양공급원이나 growth factor로 세포 성장과 활성을 증가시키는 것으로 알려져 있다 본 연구에서는 인간 진피섬유아세포를 이용하여 완두콩과 밀 펩톤이 세포증식 및 콜라겐합성에 미치는 영향과 이들의 작용기전을 규명하고자 하였다 세포증식 실험에서 완두콩과 밀 펩톤은 유의성 있게 농도 의존적으로 세포증식을 유도하였다 또한 인간 COL1A2 프로모터 루시퍼라아제와 타입 I 프로콜라겐 생합성 실험에서 완두콩 및 밀 펩톤이 COL1A2 프로모터의 활성화를 통해 타입 프로콜라겐 생합성을 촉진시키고 있음을 확인하였다 TGF-β1 루시퍼라아제 리포터 실험과 TGF-β1 ELISA 실험에서는 완두콩 및 밀 펩톤이 TGF-β1 유전자의 발현을 촉진한다는 사실을 관찰하였고 이러한 결과를 통해 완두콩 및 밀 펩톤의 콜라겐 생합성 촉진 기전이 TGF-β신호와 관련이 있음을 제시하였다 즉 완두콩 및 밀 펩톤은 TGF-β1의 발현촉진을 통해 콜라겐 생합성을 유도함을 유추할 수 있다 완두콩과 밀 펩톤을 포함한 로션을 인체피부에 주 동안 도포하여 피부자극을 관찰한 결과 어떠한 부작용도 관찰되지 않았다 이러한 연구결과를 종합해 볼 때 완두콩 및 밀펩톤이 피부자극이 없으며 피부주름을 개선시킬 수 있는 소재로 사용가능할 것으로 기대된다 Skin aging appears to be principally attributed to a decrease in both levels of Type I collagen and regeneration ability of dermal fibroblasts. It is important to introduce an efficient and safe agent for effective management of skin aging. To this end, we performed screening for anti-ageing agents and then found that vegetable peptones (pea and wheat) promoted cell proliferation of adult stem cells. Vegetable peptones may be considered as useful medium additives because it can supply nutrients, peptides, amino acids or growth factor analogues. This study was designed to investigate effects of vegetable peptones on cell proliferation/collagen production and their possible mechanisms in human dermal fibroblasts. In cell proliferation assay, vegetable peptones significantly promoted cell proliferation in a concentration-dependent manner. In addition, human COL1A2 promoter luciferase and type I procollagen synthesis assays showed that vegetable peptones induce type I procollagen production through the activation of COL1A2 promoter. In both TGF- / β1 luciferase reporter and ELISA assays, vegetable peptones was found to induce TGF- β1 production, suggesting that vegetable peptones induce type I procollagen production through the activation of TGF- β1. When applied topically in a human skin twice a day for an 4-week period of time, vegetable peptones did not induce any adverse reactions. Therefore, based on these results, we suggest the possibility that vegetable peptones may be considered as an attractive, wrinkle-reducing candidate for topical application.
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.
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.
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.