• Title/Summary/Keyword: Human Fibroblast

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Characterization Analysis for Cardiogenic Potential of Three Human Adult Stem Cells (세 종류 줄기세포의 특성 분석과 지방유래 줄기세포의 심근세포로의 분화)

  • Park, Se-Ah;Kang, Hyeon-Mi;Kim, Eun-Su;Kim, Jin-Young;Kim, Hae-Kwon
    • Development and Reproduction
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    • v.11 no.3
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    • pp.167-177
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    • 2007
  • In the present study, we isolated three human adult stem cells including adipose tissue-derived stem cells(HAD), umbilical cord-derived stem cells(HUC), and amnion-derived stem cells(HAM) and analysed their characteristics. And we examined whether HAD could be used as therapeutical cells for the heart diseases. Both HAM and HUC appeared very similar morphology but HAD was different. Doubling time of HUC was most fast, but total doubling numbers of HUC was same with HAM. Total doubling numbers of HAD was much more than others. Expression patterns of genes and proteins of three human adult stem cells were very similar. Also they were differentiated into adipocytes, osteocytes, and chondrocytes. In addition, they expressed many cardiomyocyte-related genes. But expression pattern of genes is a little different. When HAD were cultivated in the presence or absence of various combinations of BMP and FGF after 5-azacytidine expose for 24 h, expression of Cmlc-1, and ${\alpha}1c$ genes was significantly increased. However, expression of troponin T, troponin I and Kv4.3 genes was not changed. Based on these observations, it is suggested that HAD, HUC, and HAM might be used as potentially therapeutical cells for clinical application.

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The Effects of Proinflammatory Cytokines and TGF-beta, on The Fibroblast Proliferation (Proinflammatory Cytokines과 TGF-beta가 섬유모세포의 증식에 미치는 영향)

  • Kim, Chul;Park, Choon-Sik;Kim, Mi-Ho;Chang, Hun-Soo;Chung, Il-Yup;Ki, Shin-Young;Uh, Soo-Taek;Moon, Seung-Hyuk;Kim, Yong-Hoon;Lee, Hi-Bal
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.861-869
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    • 1998
  • Backgrounds: The injury of a tissue results in the infalmmation, and the imflammed tissue is replaced by the normal parenchymal cells during the process of repair. But, constitutional or repetitive damage of a tissue causes the deposition of collagen resulting in the loss of its function. These lesions are found in the lung of patients with idiopathic pulmonary fibrosis, complicated fibrosis after diffuse alveolar damage (DAD) and inorganic dust-induced lung fibrosis. The tissue from lungs of patients undergoing episodes of active and/or end-stage pulmonary fibrosis shows the accumulation of inflammatory cells, such as mononuclear cells, neutrophils, mast cells and eosinophils, and fibroblast hyperplasia. In this regard, it appears that the inflammation triggers fibroblast activation and proliferation with enhanced matrix synthesis, stimulated by inflammatory mediators such as interleukin-1 (IL-1) and/or tumor necrosis factor (TNF). It has been well known that TGF-$\beta$ enhance the proliferation of fibroblasts and the production of collagen and fibronectin, and inhibit the degradation of collagen. In this regard, It is likely that TGF-$\beta$ undergoes important roles in the pathogenesis of pulmonary fibrosis. Nevertheless, this single cytokine is not the sole regulator of the pulmonary fibrotic response. It is likely that the balance of many cytokines including TGF-$\beta$, IL-1, IL-6 and TNF-$\alpha$ regulates the pathogenesis of pulmonary fibrosis. In this study, we investigate the interaction of TGF-$\beta$, IL-1$\beta$, IL-6 and TNF-$\alpha$ and their effect on the proliferation of fibroblasts. Methods: We used a human fibroblast cell line, MRC-5 (ATCC). The culture of MRC-5 was confirmed by immunofluorecent staining. First, we determined the concentration of serum in cuture medium, in which the proliferation of MRC-5 is supressed but the survival of MRC-5 is retained. Second, we measured optical density after staining the cytokine-stimulated cells with 0.5% naphthol blue black in order to detect the effect of cytokines on the proliferation of MRC-5. Result: In the medium containing 0.5% fetal calf serum, the proliferation of MRC-5 increased by 50%, and it was maintained for 6 days. IL-1$\beta$, TNF-$\alpha$ and IL-6 induced the proliferation of MRC-5 by 45%, 160% and 120%, respectively. IL-1$\beta$ and TNF-$\alpha$ enhanced TGF-$\beta$-induced proliferation of MRC-5 by 64% and 159%, but IL-6 did not affect the TGF-$\beta$-induced proliferation. And lNF-$\alpha$-induced proliferation of MRC-5 was reduced by IL-1$\beta$ in 50%. TGF-$\beta$, TNF-$\alpha$ and both induced the proliferation of MRC-5 to 89%, 135% and 222%, respectively. Conclusions: TNF-$\alpha$, TGF-$\beta$ and IL-1$\beta$, in the order of the effectiveness, showed the induction of MRC-5 proliferation of MRC-5. TNF-$\alpha$ and IL-1$\beta$ enhance the TGF-$\beta$-induced proliferation of MRC-5, but IL-6 did not have any effect TNF-$\alpha$-induced proliferation of MRC-5 is diminished by IL-1, and TNF-$\alpha$ and TGF-$\beta$ showed a additive effect.

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Development of Monoclonal Antibody to Cytomegalovirus Maior Immediate-early(α) Protein (거대세포바이러스 조기항원 단백질에 대한 단세포군 항체 제조에 관한 연구)

  • Park, Ji Ho;Sohn, Young Mo;Park, Kyu Hyun;Lee, Won Young
    • Pediatric Infection and Vaccine
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    • v.6 no.2
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    • pp.253-260
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    • 1999
  • Purpose : The purpose of this study is to make and use monoclonal Ab which reacts with CMV major immediate early(${\alpha}$) protein(p72). Methods : Normal human fibroblast(Foreskin derived) was cultured in Eagle's minimal essential medium(MEM) containing 10% cowfetus serum and mouse chondroblast was cultured in P3X63 Ag8.653(ATCC. Maryland USA) to maintain $5{\times}10^5/ml$ cell counts. CMV(KJHJ90) from congenital CMV infected infant's urine was multiplied and used for Ab making. CMV Ag was injected 4 times, 1 week interval into the peritoneal space of 6~8 weeks old mice. And then lymphocyles and fibroblasts taken from spleen were obtained and conjugated. After the conjugated cell cultured, we chose the cell that had high Ab titer using indirect immunofluerescent method. Results : Among the 28 monoclonal antibodies obtained LPC12 and LPC23 reacted highly with nucleus of AD169 infected cell. Purified AD169 after SDS-PAGE, molecular weight of Ag, which reacted with purified monoclonal Ab, was obtained using Western blotting. Monoclonal Ab of LPC12 and LPC23 clone reacted most highly with 72 kd Ag. Conclusion : LPC12 and LPC23 clonal Ab with AD 169(P63-27) is useful on early diagnosis of CMV infection.

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Anti-aging Effects of Marine Natural Extracts against UVB-induced Damages in Human Skin Cells (UVB로 손상이 유도된 피부세포에 해양소재 추출물의 항노화 효능)

  • Lee, Chan;Jang, Jung-Hee;Kim, Bo-Ae;Park, Chan-Ik
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.38 no.3
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    • pp.255-261
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    • 2012
  • The skin is continuously exposed to environmental stresses. One of the most important stress factor is UV radiation. UV radiation causes a variety of biological effects on the skin, including inflammation, pigmentation, photoaging and cancer. Therefore in this study, we tried to search for skin-protective antioxidant materials from marine natural products (Porphyra Thalli, Laminariae japonicae thallus, Ostreae Concha, Sargassum Thallus, Undaria thallus, Haliotidis Concha, Agar, Codium thalli, Hizikia fusiforme thalli; HFE, Thalli) which exhibit protective activities against UVB-induced cytotoxicity and oxidative cell death and antiaging effects. As a results, UVB-induced cytotoxicity and cell death were effectively suppressed by treatment of Sargassum Thallus, Agar, Haliotidis Concha, Codium thalli, Thalli ethanol extracts. UVB-induced cell death was mediated by intracellular accumulation or ROS, which was significantly inhibited by treatment with marine natural products extracts. Also, The protective effect of these marine natural products seemed to be mediated by increased expression of type I collagen and Type I procollagen. These results suggest that marine natural products may have anti-aging effects new functional materials against oxidative stress-mediated skin damages.

THE CYTOTOXICITY OF PULPOTOMY MEDICAMENTS ON HUMAN PULP FIBROBLAST CELLS (치수 절단술 약제의 치수 섬유모세포에 대한 세포독성 연구)

  • Lee, Yeong-Hee;Lee, Keung-Ho
    • Journal of the korean academy of Pediatric Dentistry
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    • v.27 no.1
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    • pp.151-160
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    • 2000
  • The effect of concentration as factor in cytotoxicity, protein synthesis and alkaline phosphatase activity was compared for pulpotomy medicaments (formaldehyde, formocresol, paraformaldehyde, ferric sulfate) Human pulp fibroblasts were exposed to a range of concentrations(0.01, 0.05, 0.10, 0.25, 0.50, $1.00{\mu}l/ml$) of each agents, for period of 24hrs. The cell activities were evaluated by MTT assay, protein assay and alkaline phosphatase activity examination. The results as follows : 1. After 24hrs culture, pulp fibroblasts adding formaldehyde, formocresol and paraformaldehyde were suppressed cell activities with concentration increasing, but, no depression of cell activities by ferric sulfate. No significant difference was in formaldehyde, formocresol and paraformaldehyde. 2. Protein synthesis by pulpotomy agents were not significant difference in pulp fibroblasts but protein synthesis were a little decreased by paraformaldehyde. 3. Alkaline phosphatase activity was a little decreased by pulpotomy medicaments.

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Effects of Chlorophyll a on UVB-induced Cellular Responses and Type I pN Collagen Synthesis in vitro (클로로필 a가 UVB 유도성 산화적 스트레스와 matrix metalloproteinases (MMPs) 활성화 및 콜라겐 합성에 미치는 영향)

  • Jeon, Hee-Young;Kim, Jeong-Kee;Seo, Dae-Bang;Lee, Sang-Jun
    • Korean Journal of Food Science and Technology
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    • v.41 no.6
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    • pp.700-705
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    • 2009
  • Several studies have demonstrated that chlorophyll has many beneficial properties, including anti-inflammatory, anti-tumor, and antioxidant properties. Chlorophyll has been also shown to have an excellent chemopreventive potential against skin tumor. Its preventive mechanism against skin tumor, however, has not been examined in detail. Furthermore, the effects of chlorophyll on UVB-induced cellular responses have not been investigated. Solar UV radiation, in particular its UVB component, is the primary cause of many adverse biological effects, which is responsible for the photoaging and skin cancer. We investigated the preventive effects of chlorophyll a on UVB-mediated responses in human immortalized HaCaT kerationocytes and normal human fibroblast. We found that treatment of chlorophyll a markedly inhibited UVB-induced generation of reactive oxygen species and lipid peroxidation. Chlorophyll a also prevented UVB-induced MMP-1 expression and MMP-2 activation and increased Type I pN collagen synthesis. These results suggest that chlorophyll a is a potent candidate for the prevention and treatment of UVB-induced skin cancer and photoaging.

Anti-Wrinkle Effects of Korean Rice Wine Cake on Human Fibroblast (양조 부산물인 주박의 주름개선 효과)

  • Yoo, Jung-Min;Kang, Yeo-Jin;Pyo, Hyeong-Bae;Choung, Eui-Su;Park, Shin-Young;Choi, Ji-Ho;Han, Gwi-Jung;Lee, Choong-Hwan;Kim, Tack-Joong
    • Journal of Life Science
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    • v.20 no.12
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    • pp.1838-1843
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    • 2010
  • Skin aging is related to genetic and environmental factors (e.g., gene mutation and UV radiation respectively). To develop a new anti-wrinkle cosmetic or functional food by using Korean rice wine cake, we examined the effects of Korean rice wine cake, a brewery byproduct, on antioxidant effect, collagen synthesis and expression of MMP-1. Interestingly, we found that Korean rice wine cake has the ability to promote scavenging activity of DPPH radical. We also found that the cell proliferation and synthesis of collagen in HS27 cells was increased by Korean rice wine cake in a concentration-dependent manner. However, elastase inhibitory activity was not changed. In addition, the expression of MMP-1 was inhibited by Korean rice wine cake in a concentration-dependent manner. All these results suggest that Korean rice wine cake can be effectively used for the prevention of wrinkles in human skin.

Whitening Effect and Skin Regeneration Effect of Red Sea Cucumber Extract (홍해삼 추출물의 멜라닌 형성 억제를 통한 미백효과 및 피부 재생효과에 관한 연구)

  • Jeon, Mi Ji;Kim, Eun Ji;Kim, Geun Tae;Kim, Ga Yeon;Lee, Seung Jae;Jung, In Cheol;Kim, Sang-Yong;Kim, Young Min
    • Journal of Life Science
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    • v.28 no.6
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    • pp.681-687
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    • 2018
  • Recently, several researchers have been developing cosmetics from natural ingredients for skin whitening and anti-aging products. The red sea cucumber (RSC), Apostichopus japonicas, is a species of sea cucumber in the family stichopodiae, which is widely distributed in China, Japan, and Korea. To use Red Sea Cucumber as a cosmetic ingredient, its inhibitory effects on melanogenesis and the anti-aging effects of RSC extracts were investigated. First, a tyrosinase activity assay was performed, which showed that RSC inhibited tyrosinase activity at a concentration of $200{\mu}g/ml$. An MTT assay was carried out to evaluate cell toxicity, and the results showed that RSC extract has no cytotoxicity in HaCaT cells. Furthermore, the mRNA expression levels of tyrosinase, tyrosinase related protein 1 (TRP-1), tyrosinase related protein 2 (TRP-2), microphthalmia-associated transcription factor (MITF), and matrix metalloproteinase (MMPs) genes treated with RSC extract in B16F10 and HaCaT cells decreased. Moreover, a wound-healing assay was performed to identify the cell regeneration effect of RSC extracts. Also, a skin turnover effect was confirmed by creating a three-dimensional cell culture with HaCaT and human fibroblasts. Altogether, the results suggested that Red Sea Cucumber may possess a high ability to induce whitening and anti-wrinkle effects as a cosmeceutical ingredient.

Inhibitory Effect of Spermidine with Antioxidant Activity on Oxidative Stress in Human Dermal Fibroblasts (사람피부섬유아세포에서 산화적 스트레스에 대한 항산화 활성을 가진 spermidine의 억제효과)

  • Park, In-Hwan;Kim, Moon-Moo
    • Journal of Life Science
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    • v.21 no.5
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    • pp.693-699
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    • 2011
  • Spermidine is a ubiquitous polycation that is synthesized from putrescine, which serves as a precursor of spermine. In recent years, spermidine was found to be a polyamine that plays an important role in longevity. Reactive oxygen species (ROS) such as hydroxyl radical, superoxide and hydrogen peroxide have been shown to be involved in various pathogenic processes as well as aging. The direct scavenging effect of spermidine on DPPH radical, $H_2O_2$ and hydroxyl radical, and its protective effect against DNA oxidation related to oxidative stress were evaluated in vitro. It was observed that spermidine exhibits scavenging activities on DPPH radical and H2O2 above 500 ${\mu}M$. Spermidine was especially effective in exerting a scavenging activity on hydroxyl radical. In addition, spermidine at 1000 ${\mu}M$ showed a clear protective effect against DNA oxidation. Furthermore, the expression level of anti-oxidant enzymes such as superoxide dismutase in humam dermal fibroblasts increased in the presence of spermidine compared with blank group. These results suggest that spermidine can be used as an antioxidant to prevent ROS-related diseases including inflammation, cancer and aging.

The Effects of Orally Administered Fermented Porcine Placenta and Its Major Dipeptides on UVB-induced Wrinkle Formation in the Hairless Mice (발효 돈태반과 그 주요 다이펩타이드 섭취가 UVB 조사에 의한 무모 생쥐의 피부 주름생성에 미치는 효과)

  • Lee, JiEun;Park, Jongil;Kim, Jongbae;Jeong, Hyein;Hwang, JaeSung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.19 no.3
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    • pp.229-242
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    • 2018
  • The effects of orally administered fermented porcine placenta (FPP) and its major dipeptides, L-Leucyl-Glycine (Leu-Gly) and Glycyl-L-Leucine (Gly-Leu), on UVB-induced wrinkle formation of the skin in hairless mice was studied. Treatment with FPP, Leu-Gly or Gly-Leu increased type I procollagen synthesis and decreased MMP-1 (matrix metalloproteinase-1) in human dermal fibroblast cells (HDF-N). Hairless mice were also exposed UVB irradiation three times a week and fermented porcine placenta extract (FPP), Leu-Gly and Gly-Leu was administered once a day for eight weeks. Daily intake of FPP, Leu-Gly and Gly-Leu for eight weeks decreased wrinkles, erythema and thickness of the skin and increased skin hydration and synthesis of collagen relative to a UVB-control. Moreover, FPP, Leu-Gly or Gly-Leu intake decreased the expression of MMP-3 and MMP-13 mRNA levels and inhibited activation of MMP-2 and MMP-9 induced by UVB irradiation in hairless mice skin. These results suggest that major dipeptides of the placenta, Leu-Gly and Gly-Leu have the potential for use as a functional food ingredient with anti-wrinkling properties.