• 제목/요약/키워드: Fibroblasts, animal study

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인조혈관재료 표면에 도포된 혈관내피세포의 생리적 변화에 관한 연구 (Physiological Function of Endothelial Cells Cultured on Polyurethsne Coated by ECM)

  • 이윤신;김용배
    • 대한의용생체공학회:의공학회지
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    • 제17권2호
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    • pp.255-262
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    • 1996
  • Antithrombogenic surFace is one of the most important things to the artificial vascular prostheses. This problem will be solved if the surface of prosthesis is covered with endothelial cells. The attachment and the growth of endothelial cells onto vascular prosthesis are very difficult. So many studies have been concentrated on the attachement of endothelial cell. But no good performance of the in uiwo experiments has been shown until now. In this study, we used the whole extracellular matrix (ECM) excreted from fibroblasts as an underlying matrix, and the endothelial cells were seeded to obtain the long term patency of vascular graft(i.e., for the patent 8 week implanted wafts in the animal model of rat). In order to study the antithrombogenic functions of cultured endothelial cells, prostaglandin(PGF 1 a) synthesis and platelet adhesion were assayed. The concentration of PGF a of stimulated group was sisnificantly higher than that of control group(21.97 $\pm$ 3.45 vs 4.93 $\pm$0.71 pg/1000 cells). The platelet adhesion of the polyurethane sheet covered with endothelial cells was lower than that of polyurethane sheet or sheet covered with ECM(1.04$\pm$0.28, 2.87$\pm$0.77, 2.89$\pm$0.70, % radioactivities, respectively). Endothelial cells grew well on polyurethane coated with ECM, synthesized the prostacyclin and functioned well as antithrombogenic. Therefore the endothelialization onto the ECM excreted from fibroblasts may be a good method for the vfudig prosthesis.

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태아 섬유아세포로 핵치환된 소 난자의 핵의 재구성과정과 체외 배발달 (Nuclear Remodeling and In Vitro Development of Bovine Oocytes Following Nuclear Transfer of Bovine Fetal Fibroblasts)

  • Um, J. H.;S. J. Uhm;Kim, N-H;Lee, H. T.;K. S. Chung
    • 한국가축번식학회지
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    • 제24권1호
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    • pp.59-67
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    • 2000
  • 본 연구는 핵이 제거된 소난자 내에 소의 태아 섬유아세포를 이식한 후 난자의 발달능력을 조사하였다. 소 태아 섬유아세포는 45일된 응성 태아로부터 분리한후 MitoTracker로 염색하고 세포 주기를 동기화시키지 않은 세포를 핵이 제거된 소난자의 위란강 내에 이식하였다. 섬유아세포와 소난자의 복합체는 전기 자극을 주어 융합시키는 방법을 이용하였으며, 융합된 난자는 Calcium ionophore와 6-DMAP를 이용하여 난활성을 유도시킨 다음 CR1aa 에서 배양하였다. 핵치환 된 난자의 핵은 핵질 응축과정, 팽대과정, 전핵 형성과정이 일어났으며, 이러한 과정에서 재구성된 난자는 분열 과정을 거쳐 18∼26시간 사이에는 2-세포기로 발달하였다. 섬유아세포의 미토콘드리아는 핵치환시 난자 내로 이전되었는데 이것들은 난자 내에서 빠르게 사라지는 것으로 관찰되었으며, 핵융합 후 8시간째에는 섬유아세포의 미토콘드리아가 전혀 관찰되지 않았다. 2-세포기로 분열된 난자의 21% 가 이식 가능한 단계인 배반포 단계까지 발달하였다. 이러한 결과는 소의 태아 섬유아세포가 탈핵된 소난자 내에서 성공적으로 재분화과정을 거치며, 이렇게 재조합 된 수정란은 배반포 단계까지 발달이 가능하다는 것을 보여주고 있다.

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Antibacterial and anti-obesity effects of Abeliophyllum distichum Nakai: an in vitro study

  • Song, Dong Cheol;Lee, Ji Hwan;Oh, Han Jin;Kim, Yong Ju;An, Jae Woo;Chang, Se Yeon;Go, Young Bin;Cho, Hyun Ah;Cho, Jin Ho
    • 농업과학연구
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    • 제48권3호
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    • pp.557-565
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    • 2021
  • Interest in research on various medicinal plants has increased globally over the last few decades, possibly due to their possible antibacterial and antioxidant activities. The present study was conducted to verify the antioxidant effects, antibacterial activity, and collagen synthesis and cell viability outcomes of adipocytes upon exposure to Abeliophyllum distichum Nakai (AdN). Antibacterial activity was measured through the Disc diffusion method to compare the growth ability of pathogenic microorganisms (E.coli, Salmonella). The absorbance was measured at 560 nm to calculate the active oxygen scavenging ability. Fibroblasts were dispensed in a 96-well plate at a density of 1 × 105 cells·well-1. The amount of procollagen was measured in each case using a procollagen type 1 C-peptide EIA KIT. The cytotoxicity of the Abeliophyllum distichum Nakai extract against animal adipocytes (Hanwoo backfat cells) was determined using a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay, a method that measures the conversion of MTS to Formazan by means of mitochondrial dehydrogenases. The concentrations of the samples were made to be 0.0125, 0025, 0.05, 0.1, and 0.5% and all were -completely absorbed into the disc in an incubator at 37℃ for 24 to 36 hours. For the 0.125 mg·disc-1, effects of Abeliophyllum distichum Nakai on the antioxidant effect, antibacterial activity, and cell viability of adipocytes were found. However, Abeliophyllum distichum Nakai had no effect on collagen synthesis, thus suggesting that AdN extracts may be useful for the prevention and/or treatment of obesity.

Evaluation of Cytotoxicity for Immunity Rejection of US11, hDAF and FasL Transgene-Transfected Cells

  • Kang, Jung Won;Shin, Hyeon Yeong;Oqani, Reza K.;Lin, Tao;Lee, Jae Eun;Kim, So Yeon;Lee, Joo Bin;Jin, Dong Il
    • Reproductive and Developmental Biology
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    • 제41권3호
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    • pp.57-63
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    • 2017
  • Xenotransplantation is proposed as a solution to the problem of organ shortage. However, transplantation of xenogeneic organs induces an antigen-antibody reaction in ${\alpha}$-1,3-gal structure that are not present in humans and primates, and thus complement is also activated and organs die within minutes or hours. In this study, we used FasL gene, which is involved in the immune response of NK cell, and US11, which suppresses MHC Class I cell membrane surface expression, to inhibit cell mediated rejection in the interspecific immunity rejection, and also hDAF(CD55) was introduced to confirm the response to C3 complement. These genes were tranfeced into Korean native pig fetal fibroblasts using pCAGGS vector. And cytotoxicity of NK cell and human complement was confirmed in each cell line. The US11 inhibited the cytotoxicity of NK cell and, in addition, the simultaneous expression of US11 and Fas ligand showed excellent suppress to T-lymphocyte cytotoxicity, hDAF showed weak resistance to cytotoxicity of natural killer cell but not in CD8+ CTLs. Cytotoxicity study with human complement showed that hDAF was effective for reducing complement reaction. In this studies have demonstrated that each gene is effective in reducing immune rejection.

인체 피부 섬유아세포에서 자외선 조사에 대한 다양한 곡류 메탄올 추출물의 보호 효과 (Protective Effect of Various Grain Methanolic Extracts against UVB-induced Photo-aging in Human Skin Fibroblasts)

  • 전지영;양진우;성지혜;성예지;정헌상;이준수
    • 한국식품영양학회지
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    • 제30권2호
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    • pp.211-217
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    • 2017
  • 본 연구는 국내에서 주로 소비되는 곡류를 사용하여 UVB를 조사한 인체 피부 섬유아세포에서 항산화 효능과 MMP-1 및 수용성 콜라겐의 변화를 측정하여 광노화 억제 효능을 평가하였다. 또한 곡류 추출물의 항산화 활성 및 항산화 성분을 측정하고, 항산화력과 항산화 성분과의 상관성을 비교 분석하였다. 7종의 곡류 추출물(백미, 현미, 보리, 율무, 귀리, 수수, 흑미)은 $30mJ/cm^2$ UVB에 노출된 인체 피부 섬유아세포에서 광노화에 대해 세포 보호효과가 있는 것을 확인하였다. 또한 곡류 추출물의 처리는 UVB 조사에 따른 MMP-1의 생성을 감소시키고, 수용성 콜라겐의 분해를 저해하였으며, ROS의 생성도 억제하였다. 7종의 곡류 메탄올 추출물에 대한 총 폴리페놀 함량을 분석한 결과, 흑미와 수수가 다른 곡류에 비해 높은 항산화 성분을 함유하고 있었으며, DPPH 및 ABTS 라디칼 소거능도 높은 활성을 나타냈다. 곡류 추출물의 항산화 성분과 항산화 활성과의 상관분석 결과, 폴리페놀 함량이 높을수록 DPPH 라디칼 소거능($R^2=0.9936$), ABTS 라디칼 소거능($R^2=0.9456$)이 증가하는 것으로 나타났다. 따라서 곡류 추출물은 항산화 성분을 다량 함유하고 높은 항산화 활성을 지녔으며, UVB 조사에 의한 ROS로부터 보호하여 MMP-1의 생성을 감소시키고, 수용성 콜라겐의 분해를 억제시켜 광노화 보효호과를 나타내는 것을 확인하였다. 본 연구 결과, 곡류 추출믈의 항산화 및 광노화 보호효능 입증으로 곡류 추출물을 항주름 기능성 소재에 활용할 수 있을 것으로 판단된다.

In Vitro Development of Interspecies Nuclear Transfer Embryos: Effects of Culture Systems

  • Roh Sangho
    • Reproductive and Developmental Biology
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    • 제28권4호
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    • pp.253-256
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    • 2004
  • Porcine fibroblasts were transferred into enucleated bovine oocytes for the interspecies nuclear transfer (NT). After NT, the embryos were cultured in three different culture systems. The media used for the experiment were CR1aa and NCSU23. The culture systems used for the experiment were: 1. Culture in CR1aa for 7 days (CR). 2. Culture in CR1aa for 2 days and subsequently in NCSU23 for 5 days (CR-NC). 3. Culture in NCSU23 for 7 days (NC). Bovine (intraspecies) NT group was used as a control. The oocytes in bovine NT group were treated the same as interspecies NT embryos except using bovine fibroblasts as nuclear donors. Regardless of their nuclear origin (interspecies vs bovine), the embryos in CR (68.4% vs 77.2%) and CR-NC (67.8% vs 70.5%) showed better developmental competence to the 2-cell stage (p<0.05) than those in NC (41.0% vs 10.0%). Bovine NT embryos in CR-NC did not develop over the 4-cell stage after the medium replacement, while interspecies NT embryos in CR-NC continued to develop and could reach over the 8-cell stage (12.2%). Blastocysts were only found in bovine NT group (17.4%), but no blastocyst was found in interspecies NT group. This study suggests that the development of interspecies NT embryos mostly depends on their recipient cytoplasm during the culture in vitro.

Cytokines Regulate the Expression of the Thymus and Activation-Regulated Chemokine (TARC; CCL17) in Human Skin Fibroblast Cells

  • Lee, Ji-Sook;Kim, In-Sik;Kim, Dong-Hee;Yun, Chi-Young
    • Animal cells and systems
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    • 제10권1호
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    • pp.15-20
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    • 2006
  • Allergic inflammation is thought to be a Th2 cell-dominant immune response during which tissue-resident fibroblasts produce chemokines which contribute to the recruitment of migratory leukocytes to sites of tissue injury. Thymus and activation-regulated chemokine (TARC; CCL17) is a potent member of the CC chemokine family and a selective chemoattractant for Th2 cells. In order to study the regulatory profiles of TARC production by $TNF-{\alpha}$, $IFN-{\gamma}$, and Il-4 in human normal skin fibroblast, CCD-986sk cell line was used. The expression of TARC protein was measured using ELISA, and mRNA level was detected by RT-PCR. The combination of $TNF-{\alpha}$ and IL-4 induced a time-and dose-dependent synergistic increase in the expression of TARC at both protein and mRNA levels in the cultured human skin fibroblasts. Exposure of the cells to single cytokine had no effect on TARC expression. The high concentration (100 ng/ml) and long incubation time (72 h) of $IFN-{\gamma}$ further enhanced the TARC production induced by $TNF-{\alpha}$/lL-4 in the skin fibroblast. This synergistic effect of Th1 and Th2 type cytokines on TARC production by skin fibroblasts may contribute to the inflammatory cell infiltration and tissue damage with allergic inflammation.

Depilatory creams increase the number of hair follicles, and dermal fibroblasts expressing interleukin-6, tumor necrosis factor-α, and tumor necrosis factor-β in mouse skin

  • Tsai, Pi-Fen;Chou, Fen-Pi;Yu, Ting-Shuan;Lee, Huei-Jane;Chiu, Chun-Tang
    • The Korean Journal of Physiology and Pharmacology
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    • 제25권6호
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    • pp.497-506
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    • 2021
  • Besides using for hair removal, depilatory agents have been considered to be used as a penetration enhancer for transepidermal drug delivery. To examine the effect in hair follicles (HFs), two commercially available depilatory creams were tested on the dorsal skin of mice to monitor the effect deep into the skin structure. Fifteen male BALB/c mice were used in this study. Depilatory creams were applied to the dorsal skin of the same animal using shaved and untouched treatments as controls to minimize individual differences. Skin samples were collected at three days, one week and two weeks (n = 5 for each) after the treatment, and subjected for hematoxylin-eosin staining, and immunohistochemical analysis for proinflammatory cytokines. The morphological examination showed an increase in the thickness of epidermal layer of the depilatory cream-treated skin at early time points and in the subcutis at two weeks. Depilatory cream promoted entry of anagen phase and increased the number of hair follicles in the subcutis at one and two weeks. Immunohistochemistry showed elevated percentages of dermal fibroblasts expressing interleukin-6, tumor necrosis factor-α, and tumor necrosis factor-β. Shaving process increased the thickness of epidermis and dermis as depilatory creams did, but did neither induce the expression of proinflammatory cytokines in the dermal fibroblasts nor the number of HFs. The results suggested that the commercially available depilatory creams caused a transient minor inflammatory response of the skin and increased the levels of cytokines that might subsequently affect hair growth.

Interaction between Leptospiral Lipopolysaccharide and Toll-like Receptor 2 in Pig Fibroblast Cell Line, and Inhibitory Effect of Antibody against Leptospiral Lipopolysaccharide on Interaction

  • Guo, Yijie;Fukuda, Tomokazu;Nakamura, Shuichi;Bai, Lanlan;Xu, Jun;Kuroda, Kengo;Tomioka, Rintaro;Yoneyama, Hiroshi;Isogai, Emiko
    • Asian-Australasian Journal of Animal Sciences
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    • 제28권2호
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    • pp.273-279
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    • 2015
  • Leptospiral lipopolysaccharide (L-LPS) has shown potency in activating toll-like receptor 2 (TLR2) in pig fibroblasts (PEFs_NCC1), and causes the expression of proinflammatory cytokines. However, the stimulation by L-LPS was weak eliciting the function of TLR2 sufficiently in pig innate immunity responses during Leptospira infection. In this study, the immune response of pig embryonic fibroblast cell line (PEFs_SV40) was investigated and was found to be the high immune response, thus TLR2 is the predominate receptor of L-LPS in pig cells. Further, we found a strategy using the antibody against L-LPS, to prevent L-LPS interaction with TLR2 in pig cells which could impact on immune activation.

사람 골 형성 단백질 Ex vivo 유전자 치료법을 이용한 척추 유합 (Spinal Fusion Based on Ex Vivo Gene Therapy Using Recombinant Human BMP Adenoviruses)

  • 김기범;김재룡;안면환;서재성
    • Journal of Yeungnam Medical Science
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    • 제24권2호
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    • pp.262-274
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    • 2007
  • AdBMP-2와 AdBMP-7을 형질도입 시킨 사람 섬유아세포와 사람 골수기질세포를 면역결핍 생쥐의 척추 옆 근육으로 주입하여 척추 유합을 유도한 결과, AdBMP-7/BMSC가 AdBMP-2/BMSC 또는 AdBMP-7/HuFb와 AdBMP-2/HuFb 보다 골 형성능이 우수하였으며, 척추 유합을 잘 유도하였음을 확인하였다.

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