• 제목/요약/키워드: 3T3 fibroblast

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Quantitative Analysis of Growth of Cells on Physicochemically Modified Surfaces

  • Chandra, Prakash;Kim, Jihee;Rhee, Seog Woo
    • Bulletin of the Korean Chemical Society
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    • 제34권2호
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    • pp.524-530
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    • 2013
  • In this study, we describe the most expected behavior of cells on the modified surface and the correlation between the modified substrates and the response of cells. The physicochemical characteristics of substrates played an essential role in the adhesion and proliferation of cells. Glass and polymer substrates were modified using air plasma oxidation, and the surfaces were coated with self-assembled monolayer molecules of silanes. The PDMS substrates embedded with parallel micropatterns were used for evaluation of the effect of topologically modified substrate on cellular behaviour. BALB/3T3 fibroblast cells were cultured on different surfaces with distinct wettability and topology, and the growth rates and morphological change of cells were analyzed. Finally, we found the optimum conditions for the adhesion and proliferation of cells on the modified surface. This study will provide insight into the cell-surface interaction and contribute to tissue engineering applications.

유선상피 간세포의 분리 및 특성연구 (Isolation and Characterization of Mammary FpithelialStem Cells in Culture)

  • 백기주;류환선;;김남득
    • 생명과학회지
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    • 제10권1호
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    • pp.37-44
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    • 2000
  • The mammary gland contains a subpopulation of epithelial cells with large proliferative potentials which are the likely targets for carcinogens. These clonogenic cells can proliferate and differentiate into functional glandular structures. Rat mammary epithelial cells (RMEC) were isolated and characterized in vitro. By flow cytometry of RMEC stained with fluorescein isothiocyanate-peanut agglutinin(PNA) and phycoerythrin anti-Thy-1.1 monoclonal antibody, it was possible to four cell subpopulations from 7-8 week old F344 female rat mammary glands: cells negative to both reagents (B-), PNA-positive cells (PNA+), Thy-1.1-positive cells (Thy-1.1+), and cells positive to both reagents (B+). When single PNA+ cells were isolated and cultured in Matrigel with irradiated (∼50 Gray) 3T3 fibroblast feeder layer, they gave rise to multicellular clonal structures of three types: alveolar, foamy alveolar, and squamous colonies. The developed structures were similar to the mammary glands in vivo. These results suggest that some of PNA+ cells possesses many of the characteristics of multipotent clonogenic stem-like cells.

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In vitro Biodegradability and Surface Properties of Block Copoly(ester-ether)s Consisting of Poly(L-lactide) and Polyether

  • Lee, Chan-Woo;Kim, Yoshiharu ura
    • Macromolecular Research
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    • 제11권1호
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    • pp.42-46
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    • 2003
  • Cell attachment and proliferation on the polymer films of triblock copolymer(ester-ether)s comprising po1y (L-1actide) (PLLA) and poly (oxyethylene-co-oxypropylene)(PN) were investigated using 3T3 fibroblasts. It was found that on the tissue culture polystyrene(TCPS) and the PLLA control film the cells could spread well while on the copolymer films the cells showed a rounded morphology without spreading and proliferated weakly. Especially, little cells proliferated on the films of copolymer having a LN composition of 20 wt%. While the water absorption of the copolymer films increased with increasing PN content, the contact angle against water of copolymer films immersed in aqueous medium was almost identical, being slightly lower than that of the PLLA film. These properties were compatible with the results of cell attachment. The in vitro hydrolysis of the films of triblock and multiblock type copolymers was faster with increasing PN content. The increased hydrolyzability, the flexibility and the decreased cell attachment suggested that these copolymers may have high potential as biodegradable materials for medical use.

Effect of Poly(3-hydroxibutyrate-co-3-hydroxivalerate) Surface with Different Wettability on Fibroblast Behavior

  • Lee, Sang-Jin;Lee, Young-Moo;Khang, Gilson;Kim, Un-Young;Lee, Bong;Lee, Hai-Bang
    • Macromolecular Research
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    • 제10권3호
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    • pp.150-157
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    • 2002
  • Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) is a microbial storage polymer with biodegradable properties. In order to improve the cell compatibility of PHBV surfaces, the physicochemical treatments have been demonstrated. In this study, physical method was corona discharge treatment and chemical method was chloric acid mixture solution treatment. The physicochemically treated PHBV film surfaces were characterized by the measurement of water contact angle, electron spectroscopy for chemical analysis, and scanning electron microscopy (SEM). The water contact angle of the physicochemically treated PHBV surfaces decreased from 75 to 30~40 degree, increased hydrophilicity. due to the introduction of oxygen-based functional group onto the PHBV backbone chain. The mouse NIH/3T3 fibroblasts cultured onto the physicochemically treated PHBV film surfaces with different wettability. The effect of the PHBV surface with different wettability was determined by SEM as counts of cell number and [$^3$H]thymidine incorporation as measures of cell proliferation. As the surface wettability increased, the number of the cell adhered and proliferated on the surface was increased. The result seems closely related with the serum protein adsorption on the physicochemically treated PHBV surface. In conclusion, this study demonstrated that the surface wettabilily of biodegradable polymer as the PHBV plays an important role for cell adhesion and proliferation behavior for biomedical application.

어혈처방 3종이 지방전구세포 분화에 미치는 영향 (Anti-adipogenesis Effects of 3 Herbal Formula on Blood Stasis)

  • 이호영;심은형;이명수;이주아
    • 대한본초학회지
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    • 제31권6호
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    • pp.29-35
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    • 2016
  • Objectives : Blood stasis (BS) is related to be caused by blood circulation and stagnation which are cancer, atherosclerosis and hyperlipidemia in traditional medicine. We extracted 3 kinds of BS formula; Seogakjihwang-tang (SGT), Tonggyuhawlhyul-tang (THT), Hyulbuchukeo-tang (HCT). This study was conducted to investigate whether the 3 kinds of herbal formula extracts have inhibitory efficacy association with anti-adipogenesis. Methods : To investigate the anti-adipogenesis, we used the mouse fibroblast cell line, 3T3-L1 which differentiated into adipocytes in response to insulin, IBMX and dexamethasone (MDI). Cytotoxicity of herbal formula extracts were examined by CCK-8 kit. Intracellular lipid droplets were detected by Oil-Red-O staining. Triglyceride (TG) and leptin were measure using elisa kit. Results : The yield of water extracts was 14.62% (SGT), 21.27% (THT), 20.02% (HCT). Lipid accumulation was reduced significantly by 3 kinds of herbal formula compared to control. Especially, THT and HCT decreased lipid droplet, respectively at all concentration. The TG and leptin were also inhibited by 3 kinds of herbal formula. The IC50 of TG were $280.51{\mu}g/m{\ell}$ (SGT), $52.62{\mu}g/m{\ell}$ (THT), $313.99{\mu}g/m{\ell}$ (HCT). The IC50 of leptin were $348.76{\mu}g/m{\ell}$ (SGT), $164.02{\mu}g/m{\ell}$ (THT), $257.00{\mu}g/m{\ell}$ (HCT). THT was better than other herbal formula on anti-adipogenesis. Conclusion : kinds of herbal formula inhibited adipogenesis in MDI-induced 3T3-L1 adipocytes, as indicated by the significant reduction in TG and leptin concentration without cytotoxicity. Therefore, 3 kinds of herbal formula for BS might act as a therapeutic agent for preventing lipid diseases, such as obesity and atherosclerosis.

Tissue engineering of dental pulp on type I collagen

  • Lee, Gwang-Hee;Huh, Sung-Yoon;Park, Sang-Hyuk
    • Restorative Dentistry and Endodontics
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    • 제29권4호
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    • pp.370-377
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    • 2004
  • The purpose of this study was to regenerate human dental pulp tissues similar to native pulp tissues. Using the mixture of type I collagen solution, primary cells collected from the different tissues (pulp, gingiva, and skin) and NIH 3T3 ($1{\;}{\times}{\;}10^5{\;}cells/ml/well$) were cultured at 12-well plate at $37^{\circ}C$ for 14 days. Standardized photographs were taken with digital camera during 14 days and the diameter of the contracted collagen gel matrix was measured and statistically analyzed with student t-test. As one of the pulp tissue engineering, normal human dental pulp tissue and collagen gel matrix cultured with dental pulp cells for 14 days were fixed and stained with Hematoxyline & Eosin. According to this study, the results were as follows: 1. The contraction of collagen gel matrix cultured with pulp cells for 14 days was significantly higher than other fibroblasts (gingiva, skin) (p < 0.05), 2. The diameter of collagen gel matrix cultured with pulp cells was reduced to 70.4% after 7 days, and 57.1% after 14 days. 3. The collagen gel without any cells did not contract, whereas the collagen gel cultured with gingiva and skin showed mild contraction after 14 days (88.1% and 87.6% respectively). 4. The contraction of the collagen gel cultured with NIH 3T3 cells after 14 days was higher than those cultured with gingival and skin fibroblasts, but it was not statistically significant (72.1%, p > 0.05). 5. The collagen gel matrix cultured with pulp cells for 14 days showed similar shape with native pulp tissue without blood vessels. This approach may provide a means of engineering a variety of other oral tissue as well and these cell behaviors may provide information needed to establish pulp tissue engineering protocols.

Mitochondria-Targeted Vitamin E Protects Skin from UVB-Irradiation

  • Kim, Won-Serk;Kim, Ikyon;Kim, Wang-Kyun;Choi, Ju-Yeon;Kim, Doo Yeong;Moon, Sung-Guk;Min, Hyung-Keun;Song, Min-Kyu;Sung, Jong-Hyuk
    • Biomolecules & Therapeutics
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    • 제24권3호
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    • pp.305-311
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    • 2016
  • Mitochondria-targeted vitamin E (MVE) is designed to accumulate within mitochondria and is applied to decrease mitochondrial oxidative damage. However, the protective effects of MVE in skin cells have not been identified. We investigated the protective effect of MVE against UVB in dermal fibroblasts and immortalized human keratinocyte cell line (HaCaT). In addition, we studied the wound-healing effect of MVE in animal models. We found that MVE increased the proliferation and survival of fibroblasts at low concentration (i.e., nM ranges). In addition, MVE increased collagen production and downregulated matrix metalloproteinase1. MVE also increased the proliferation and survival of HaCaT cells. UVB increased reactive oxygen species (ROS) production in fibroblasts and HaCaT cells, while MVE decreased ROS production at low concentration. In an animal experiment, MVE accelerated wound healing from laser-induced skin damage. These results collectively suggest that low dose MVE protects skin from UVB irradiation. Therefore, MVE can be developed as a cosmetic raw material.

Effects of 60-Hz Magnetic Fields on DNA Damage Responses in HT22 Mouse Hippocampal Cell Lines

  • Mun, Gil-Im;Lee, Seungwoo;Kim, Nam;Lee, Yun-Sil
    • Journal of electromagnetic engineering and science
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    • 제15권3호
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    • pp.123-128
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    • 2015
  • Previously, we investigated extremely low-frequency magnetic fields (ELF-MFs) on diverse DNA damage responses, such as phosphorylated H2AX (${\gamma}H2AX$), comet tail moments, and aneuploidy production in several non-tumorigenic epithelial or fibroblast cell lines. However, the effect of ELF-MF on DNA damage responses in neuronal cells may not be well evaluated. Here, we investigated the effects of ELF-MF on the DNA damage responses in HT22 non-tumorigenic mouse neuronal cells. Exposure to a 60-Hz, 2 mT ELF-MF did not produce any increased ${\gamma}H2AX$ expression, comet tail moments, or aneuploidy formation. However, 2 mT ELF-MF transiently increased the cell number. From the results, ELF-MF could affect the DNA damage responses differently, depending on the cell lines.

혈당저하를 위해 처방된 천연물 복합제의 항산화 활성 및 3T3-L1 세포의 분화에 미치는 영향 (Anti-oxidative activity of the herb mixture prescribed to induce blood glucose level and effect on the differentiation of 3T3-L1 fibroblast)

  • 김정옥;김정배;김학윤
    • 한국식품저장유통학회지
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    • 제16권1호
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    • pp.115-121
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    • 2009
  • 혈당저하를 위한 천연물복합제를 한의학 처방 및 문헌 등을 기초로 하여 구성하고, 기능성 식품 소재로서의 이용가능성을 조사하기 위하여 추출물의 이화학적 특성을 조사하였다. 천연물복합제 5가지 추출물의 수율은 $19.52{\sim}29.79%$의 범위로 나타났으며, 총페놀성 화합물 함량은 $349.24{\sim}1752.21\;mg%$, 총 플라보노이드 함량은 $163.06{\sim}1,118.47\;mg%$의 범위로 조사되었다. 특히 2번, 3번 및 5번 복합제에서 1,000 mg% 이상의 높은 함량을 나타내었으며, 전자공여능 또한 이들 복합제에서 높은 활성을 보여주었다. 아질산염 소거능은 2번 및 5번 천연물복합제가 pH 1.2에서 70% 이상의 높은 활성을 보여주었으며, pH가 증가할 수록 감소하였다. 인슐린성 물질의 함유정도를 조사하기 위하여 섬유아세포 3T3-L1의 지방세포로의 분화정도를 측정한 결과, 천연물복합제 5가지 추출물 중 5, 1, 4, 3, 2번의 순으로 높은 값을 나타내었다. 한편 이들 천연물복합제의 기능성 식품으로 이용 가능성을 검증하기 위한 추가적인 생리활성 및 효능 연구를 진행할 필요성이 있다고 판단된다.

택사추출물의 성분분리와 화장품 원료로서의 특성 (Separation and Purification of Effective Components from the Alisma orientale and its Application as a Cosmeceutical Ingredient)

  • 이대우;김영진;김영실;김종헌
    • 대한화장품학회지
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    • 제32권1호
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    • pp.23-28
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    • 2006
  • 본 연구는 택사의 화장품 원료로서의 특성을 알아보기 위하여 화장품의 기능들인 항산화, 미백, 세포 손상 회복 및 항염증과 관련된 다양한 실험을 실시하였다. 30, 70, 100% MeOH 용매로 추출한 택사 추출물들은 DPPH법으로 실시한 free radical scavenging assay에서 좋은 활성을 보여주었고, tyrosinase 활성 억제 시험에서도 0.5% 이상의 농도에서 농도 의존적인 활성을 보여주었다. Human fibroblast를 사용한 proliferation assay (MTT assay)에서 각 용매 추출물들은 별다른 효과를 보여주지 못했고 0.05% 이하의 농도에서는 세포 독성으로부터 안전하다는 것을 알 수 있었다. Bl6 melanocyte를 사용한 melanin 생성 억제시험에서 각 용매별 추출물은 독성으로부터 안전한 0.05% 이하의 농도에서 melanin 생성을 40% 이상 억제하는 높은 활성을 보여주었다. 이후 우리는 택사 추출물의 MPLC분리 분획을 실시하여 세 가지 분획을 얻었으며 이들을 대상으로 세포 손상 회복시험과 melanin생성 억제 시험, 염증인자 생성 억제 시험을 실시하였다. 그 결과 분획물들 중 3번 분획물이 세포 손상 회복을 30% 이상 올려주는 좋은 결과를 보여주었고, melanin 생성 억제 시험과 COX-2 생성억제에서도 주목할만한 결과를 보여주었다.