• 제목/요약/키워드: aggregate culture

검색결과 47건 처리시간 0.031초

연골세포와 중간엽줄기세포의 3차원 Co-culture를 통한 연골화 향상 (Enhanced Chondrogenesis by Three-dimensional Co-culture of Chondrocytes and Mesenchymal Stem Cells)

  • 황슬기;차현명;임진혁;이지희;심혜은;김동일
    • KSBB Journal
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    • 제31권2호
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    • pp.120-125
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    • 2016
  • Two-dimensional cultivation is typically used for cell growth, but the method reduces the characteristics of chondrocytes and stem cells, and limits culture area. Therefore, development of three-dimensional culture method is needed to mimic in vivo environment, improve quality of cells and scale-up efficiently. Improving proliferation and chondrogenesis is available by co-culture of chondrocytes and mesenchymal stem cells (MSCs) that leads to interaction between two kinds of cells. However, the co-culture has problems that permeability of sphere diminishes as aggregate size increased and ratio of two kinds of cells composing each spheres is different. In this work, co-cultivation method using controlled sphere composed of chondrocytes and MSCs was established and enhanced chondrogenesis. Periosteum-derived progenitor cells (PDPCs) that are appropriate for cell therapy source of articular cartilage were used as MSCs. Controlled spheres were formed in the hanging-drop plates and shifted for being induced chondrogenesis in 35-mm non-adhesive culture dishes at a rotation rate of 60 rpm. After inducing chondrogenesis, gene expressions related with chondrogenesis were found to be improved and it was apparent that the utilization of controlled spheres promoted chondrogenesis. As a result, available numbers of cells per unit area were increased and chondrogenic differentiation ability was improved compared to typical two-dimensional culture. This approach shows the potential in cartilage regeneration as it can provide sufficient numbers of chondrocytes.

Production of Bacterial Cellulose by Gluconacetobacter hansenii PJK Isolated from Rotten Apple

  • Park, Joong-Kon;Park, Youn-Hee;Jung, Jae-Yong
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제8권2호
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    • pp.83-88
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    • 2003
  • A cellulose-producing strain isolated from rotten apples was identified as Gluconacetobacter hansenii based on its physiological properties and the 16S rDNA complete sequencing method, and specifically named Gluconacetobacter hansenii PJK. The amount of bacterial cellulose (BC) produced by G. hansenii PJK in a shaking incubator was 1.5 times higher than that produced in a static culture. The addition of ethanol to the medium during cultivation enhanced the productivity of bacterial cellulose, plus the supplementation of 1% ethanol into the culture medium made the produced BC aggregate into a big lump and thus protected the bacterial-cellulose-producing G. hansenii PJK cells in the shear stress field from being converted into non-cellulose-producing (Cel) mutants. Cells subcultured three times in a medium containing ethanol retained their ability to produce BC without any loss in the production yield.

Enhancement of anti-inflammatory and anti-tumorigenic properties of 3D-spheroid formed mesenchymal stem cells derived from rheumatoid arthritis joints

  • Seung-Chan Lee;Chae-Yeon Hong;Yong-Ho Choe;Tae-Seok Kim;Won-Jae Lee;Gyu-Jin Rho;Sung-Lim Lee
    • 한국동물생명공학회지
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    • 제37권4호
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    • pp.246-254
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    • 2022
  • Current studies have revealed the capacity of mesenchymal stem cells (MSCs) in term of immunomodulatory properties, and this distinct potential is downgraded according to the disease duration of patients-derived MSCs. In order to enhance the immunomodulatory and anti-tumorigenic properties of the rheumatoid arthritis (RA) joints-derived MSCs, we aggregate synovial fluid-derived MSCs from RA joints (RA-hMSCs) into 3D-spheroids by the use of hanging drop culture method. Cells were isolated from synovial fluids of RA joints with longstanding active status over 13 years. For aggregation of RA-hMSCs into 3D-spheroids, cells were plated in hanging drops in 30 μL of advanced DMEM (ADMEM) containing 25,000-30,000 cells/drop and cultured for 48 h. To analyze the comparative immunomodulatory effects of 3D-spheroid and 2D monolayer cultured RA-hMSCs and then cells were cultured in ADMEM supplemented with 20% of synovial fluids of RA patients for 48 h and were evaluated by qRT-PCR for their expression of mRNA levels of inflammatory and anti-inflammatory markers. Cellular aggregation of RA-hMSCs was observed and cells were aggregate into a single sphere. Following treatment of RA patient's synovial fluids into the RA-hMSCs, spheroids formed RA-hMSCs showed significantly (p < 0.05) higher expression of TNFα stimulated gene/protein 6 (TSG-6) than the monolayer cultured RA-hMSCs. Therefore, the 3D-spheroid culture methods of RA-hMSCs were more effective than 2D monolayer cultures in suppressing inflammatory response treated with 20% of RA-synovial fluids by expression of TNFα (TSG-6) according to the immune response and enhanced secretion of inflammatory factors.

Suspension Culture-Mediated Tetraploid Formation in Mouse Embryonic Stem Cells

  • Lee, Jae-Hee;Gong, Seung-Pyo;Lim, Jeong-Mook;Lee, Seung-Tae
    • Reproductive and Developmental Biology
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    • 제36권1호
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    • pp.21-26
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    • 2012
  • Suspension culture is a useful tool for culturing embryonic stem (ES) cells in large-scale, but the stability of pluripotency and karyotype has to be maintained $in$ $vitro$ for clinical application. Therefore, we investigated whether the chromosomal abnormality of ES cells was induced in suspension culture or not. The ES cells were cultured in suspension as a form of aggregate with or without mouse embryonic fibroblasts (MEFs), and 0 or 1,000 U/ml leukemia inhibitory factor (LIF) was treated to suspended ES cells. After culturing ES cells in suspension, their karyotype, DNA content, and properties of pluripotency and differentiation were evaluated. As a result, the formation of tetraploid ES cell population was significantly increased in suspension culture in which ES cells were co-cultured with both MEFs and LIF. Tetraploid ES cell population was also generated when ES cells were cultured alone in suspension regardless of the existence of LIF. On the other hand, the formation of tetraploid ES cell population was not detected in LIF-free condition, in which MEFs were included. The origin of tetraploid ES cell population was turned out to be E14 ES cells and not MEFs by microsatellite analysis and the basic properties of them were still maintained despite ploidy-conversion to tetraploidy. Furthermore, we identified the ploidy shift from tetraploidy to near-triploidy as tetraploid ES cells were differentiated spontaneously. From these results, we demonstrated that suspension culture system could induce ploidy-conversion generating tetraploid ES cell population. Moreover, optimization of suspension culture system may make possible mass-production of ES cells.

Zinc Metal Solubilization by Gluconacetobacter diazotrophicus and Induction of Pleomorphic Cells

  • Saravanan, Venkatakrishnan Sivaraj;Osborne, Jabez;Madhaiyan, Munusamy;Mathew, Lazar;Chung, Jong-Bae;Ahn, Ki-Sup;Sa, Tong-Min
    • Journal of Microbiology and Biotechnology
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    • 제17권9호
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    • pp.1477-1482
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    • 2007
  • Gluconacetobacter diazotrophicus strain PA15 exhibited a minimum inhibitory concentration value of 11 mM in an LGI medium amended with $ZnCl_2$. When an LGI medium was amended with Zn metal, solubilization halos were observed in a plate assay, and further solubilization was confirmed in a broth assay. The maximum solubilization was recorded after 120 h with a 0.1% Zn metal amendment. During solubilization, the culture growth and pH of the broth were indirectly correlated. Using a Fourier Transform Infrared Spectroscopy analysis, one of the agents solubilizing the Zn metal was identified as gluconic acid. When the Zn-amended broth was observed under a bright field microscope, long involution cells were observed, and further analysis with Atomic Force Microscopy revealed highly deformed, pleomorphic, aggregate-like cells.

산화철 나노입자의 마이크로캡슐화와 이를 이용한 세포의 자력부상 배양 (Microencapsulation of Iron Oxide Nanoparticles and Their Application in Magnetic Levitation of Cells)

  • 이진실;이준호;심재권;허원
    • 공업화학
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    • 제31권1호
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    • pp.13-18
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    • 2020
  • 실크의 섬유 고분자 단백질인 피브로인을 사용하여 산화철 나노입자가 내포된 테라그노시스가 가능한 마이크로캡슐을 제조하였다. 열중량 분석으로 산화철의 함량은 4.28%, 자력계로는 5.11%로 측정되었다. 산화철 마이크로캡슐이 첨가된 마우스 섬유아세포 3T3 배양액에서 얻어진 세포 현탁액은 자력에 반응하여 맑게 변하고, 세포는 자석 방향으로 응집하였다. 배양접시 상단에 올려둔 네오디뮴 자석은 세포를 배양액 표면 중심으로 세포를 끌어모았다. 배양액 표면에 모인 세포들은 응집하여 72 h 이후 장축의 길이가 2 mm인 비대칭 타원체인 세포 집합체를 형성하였다. 세포집합체의 바깥층에는 세포들이 상대적으로 크고 서로 모여 치밀한 조직을 형성하였으나, 중심부는 물질전달제한으로 세포의 사멸이 진행되는 것으로 관찰되었다. 바깥층에는 산화철 마이크로캡슐이 자력의 방향으로 체인처럼 일렬로 늘어선 현상도 관찰되었다. 마이크로CT를 이용하여 세포응집체 내부의 산화철이 고루 분포하지 않고 자력 방향으로 비대칭적으로 분포하고 있음을 보였다.

현대 패션에 나타난 문화적 퓨전 현상에 관한 연구 (A Study on Cultural Fusion in Modern Fashion)

  • 김소영;강경애
    • 한국의상디자인학회지
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    • 제9권2호
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    • pp.167-178
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    • 2007
  • The purpose of this study is to systematically examine fusion phenomenon in the cultural aspect aiming to grasp the characteristics of fusion phenomenon, which has a steady impact on fashion trend. A reason for fusion to be noted as a cultural code is because diversely cultural interpretations are impossible from one viewpoint. Therefore, it is aimed to inquire into the characteristics of fusion culture, which is one phenomenon of newly mass culture, and to examine which cultural fusion phenomenon in modern fashion is being displayed in fashion design. First, the culturally fusion phenomenon, which was indicated in modern fashion, is displayed as repetition. The fusion, which integrates into fashion as a cultural code, is a trend of pursuing the experience with different culture. The needs of consumers are shown as fusion fashion that repeatedly uses the traditional detail and the modern detail through cultural repetition. Second, it is displayed as transformation. A concept of fusion is not the simple aggregate in [1+1=2], but the creation of new culture called [1+1=1]. In modern fashion, the cultures in the East and the West are harmoniously transformed, and through transformation, each culture is being compressively delivered. Third, it is displayed as interdependency. The fusion fashion which was newly re-interpreted while having reciprocal influence upon mutual culture, is re-interpreted by forming the equal relationship in heterogeneous elements without destroying fashion that was made previously.

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전통공연예술 산업의 글로벌 경쟁력확보를 위한 전략적 방안 모색에 관한 연구 (Study on Strategic Plan for Ensuring the Global Competitiveness of Traditional Performing Arts Industry)

  • 임영순;맹해양;배기형
    • 한국콘텐츠학회논문지
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    • 제16권4호
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    • pp.88-99
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    • 2016
  • 전통공연예술은 한 국가의 정체성을 나타내는 국가경쟁력의 핵심요소로 많은 국가들은 자국의 독특한 전통공연예술 장르를 대중화, 산업화하는 등의 노력을 기울이고 있다. 전통예술은 문학, 미술, 음악, 무용, 연극 등 총체적인 집합체로 이러한 전통문화를 보존하고 미래의 문화에 부합하도록 활성화하는 것은 경쟁력 있는 국가브랜드를 확보하는 일이라고 볼 수 있다. 그동안 K-pop 및 드라마 등 우리 대중문화 콘텐츠의 세계적인 진출성공사례에 가려져 있지만 우리 전통공연예술분야는 꾸준한 역량 확대 및 세계의 주목을 받는 노력과 성과가 있어 왔다. 이제는 전통공연예술 분야에서 국내 및 세계에서 보다 대중적인 수요를 확대하기 위해 새로운 정책적 접근이 필요한 시점일 것이다. 따라서 본 연구에서는 전통공연예술 장르의 대중화, 산업화의 필요와 우리만의 독특한 전통공연예술 콘텐츠의 성공적인 해외 진출 및 진흥기회를 마련하기 위한 전략적 방안을 모색해 보고자 한다. 이를 위하여 먼저 전통공연예술의 개념을 정의하고 전통공연예술 산업의 현황을 정리하였다. 또한 이러한 전통공연예술 산업의 현황을 토대로 우리나라 전통공연예술 콘텐츠의 해외 시장 저변확대를 위한 전략적 방안을 제안하였다.

Mineral trioxide aggregate가 유치 및 영구치의 치수기질세포 증식 및 분화에 미치는 영향 (Effects of Mineral Trioxide Aggregate on the Proliferation and Differentiation of Human Dental Pulp Stromal Cells from Permanent and Deciduous Teeth)

  • 김승혜;전미정;신동민;이제호;송제선
    • 대한소아치과학회지
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    • 제40권3호
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    • pp.185-193
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    • 2013
  • 최근 유구치의 치수절단술 약제로 MTA의 임상 적용이 문헌들에서 보고된 바 있으나 MTA 표면에서 일어나는 유치 치수 세포의 반응에 대한 시험관내 연구는 많이 보고되지 않았다. 이번 연구의 목적은 유치 및 영구치에서 유래한 치수기질세포가 경화된 MTA 표면에서 나타내는 증식 및 분화 능력을 비교 평가하는 것이었다. 사람 영구치와 유치 치수 조직에서 분리된 치수기질세포를 경화된 MTA 표면에서 배양 후 세포증식율과 세포주기를 검사하였으며, 정량적 역전사 중합효소 연쇄반응(RT-PCR)을 사용하여 분화양상을 분석하였다. Runt-related transcription factor 2(Runx2)와 alkaline phosphatase(ALP)가 정량적 RT-PCR의 표지자로 사용되었고, MTA 표면에서 증식된 치수기질세포의 형태학적 변화를 주사전자현미경 하에서 관찰하였다. 영구치와 유치의 치수기질세포군은 세포증식률, 세포주기 분포 및 mRNA 발현 양상에 있어서 차이를 보이지 않았으며, 주사전자현미경 상에서 두 군 모두 수지상 형태를 나타내었다. MTA 상에서 관찰된 유치와 영구치의 치수기질세포의 비슷한 증식력 및 광화를 유도하는 세포로의 분화능은 유치의 치수절단술 제재로 MTA가 생체친화적으로 적합함을 보여준다.

수종의 치근단역충전 재료가 MG63 osteoblast-like cells에 미치는 영향 (THE EFFECT OF SEVERAL ROOT-END FILLING MATERIALS ON MG63 OSTEOBLAST-LIKE CELLS)

  • 이정호;손원준;이원철;백성호
    • Restorative Dentistry and Endodontics
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    • 제35권3호
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    • pp.222-228
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    • 2010
  • 본 연구의 목적은 현재 치근단 역충전재로 널리 사용되고 있는 MTA와 새롭게 개발된 제품인 DB를 MG63 세포를 사용하여 비교하는 것이다. 치근단 역충전재료로는 MTA, DB, IRM을 사용하였고 대조군으로는 tissue culture plastic을 사용하였다. 각 재료를 혼합하고 혼합물의 경화가 일어나도록 24시간 동안 놓아두었다. MG63 세포를 각 군에 뿌려준 후 세포가 재료에 부착될 수 있도록 4시간 배양하였다. 치근단 역충전재를 세포에 접촉시킨 후 세포수준의 초기 반응을 관찰하였다. 12시간 더 배양한 후 세포가 각 재료에 붙어 있는 정도를 관찰하고, 각 재료가 골형성에 미치는 영향을 알아보기 위해 ELISA를 이용하여 $TGF{\beta}1$, OC를 측정하였고 ALP의 양도 측정하였다. 결과는 일원배치분산분석법으로 통계처리하였다. 그 결과, 재료에 부착이 일어난 세포의 수 항목과 OC 항목에서만 MTA와 DB간에 통계적으로 차이가 없었다. 다른 항목들에서는 모든 군 간에 통계적으로 유의한 차이가 있었다. DB가 MTA와 완전히 같은 세포반응을 보이지는 않았지만 부착이 일어난 세포의 수는 재료의 생체적합성을 나타내며 OC는 골형성 정도를 나타내므로 DB가 역충전 재료로 사용된다면 MTA와 유사한 결과를 보일 것으로 예측된다.