• 한국가축번식학회지
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• 제27권1호
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• pp.53-59
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• 2003

본 연구에서는 한우 태아의 시기별로 35일령, 50일령, 70일령 및 90일령의 fetal fibroblast cell line을 생산하였고, bovine-specific primer와 Y chromosome-specific primer를 이용하여 PCR에 의해 성을 판별하여 각각 암수 2 line의 한우 fetal fibroblast cell line을 확립하였다. 이들 cell line을 계대배양하여 passage number가 10 이상에서 염색체 분석을 실시하였는데 모두에서 80%이상의 세포가 60개의 정상 염색체수의 나타내어 계대배양이 karyotype에 영향을 미치지 않는 것으로 나타났다. Serum starvation과 confluent 배양 방법을 이용하여 Go 상태로 유도되었는지 확인하기 위해 PCNA antibody를 이용하여 Western blotting 분석을 실시하였는데 PCNA 발현이 현저히 감소되는 것을 확인할 수 있었고, 다시 정상 medium으로 환원시켰을 때 세포분열이 재개되어 Go상태로 유도되었음을 확인할 수 있었다. 또한 serum stravation 방법이 conflent한 배양방법보다 PCNA 발현양이 적은 것으로 나타나 좀더 효율적인 Go 상태 세포 주기 조절방법으로 판명되었다.

• Journal of Periodontal and Implant Science
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• 제25권1호
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• pp.133-145
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• 1995

The migration and proliferation of periodontal ligament cells are desired goal of periodontal regeneration therapy. PDGF and $TGF-{\beta}1$ are well known to regulate the cell activity of mesenchymal origin cell. The purpose of this study was to determine the effects of these growth factors on human gingival fibroblast and periodontal ligament cell actvity, and to identify the regulatory effect of $TGF-{\beta}1$ on the response to PDGF by MIT assay. Human gingival fibroblast and periodontal ligament cells were cultured from extracted teeth for non-periodontal reason. Cultured human gingival fibroblast and periodontal ligament cells in vitro were treated with polyperpetide growth factor PDGF and $TGF-{\beta}1$ in both a dose and time - dependent manner. Cell morphology were determined by inverted microscope and cell acitivity were determined by MIT assay. The result of this study demonstrated that PDGF and $TGF-{\beta}1$ were not changed the morphology of these cell compared with control group. PDGF or $TGF-{\beta}1$ increased cell activity of periodontal ligament cell in dose and time dependent manner but gingival fibroblast were decreased to the level of control group at third day. Additionally, incubation with $TGF-{\beta}1$ addition to PDGF resulted in a enhanced cell activity of PDGF. Therefore, cell acitivty of gingival fibroblast were not changed compared with control group. This stiudy demonstrates that PDGF and $TGF-{\beta}1$ are major mitogens for human periodontal ligament cell in vitro, and $TGF-{\beta}1$ is a regulator of cell activity to PDGF in human gingival fibroblast and periodontal ligament cell.

• 대한한방부인과학회지
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• 제24권2호
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• pp.13-32
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• 2011

Objectives: This study was performed to elucidate the effects of Danchisoyo-san (DS) on cell damage and gene expression in UVB-exposed dermal fibroblast. Methods: To demonstrate the inhibitory effects of DS on aging of the skin, we used human dermal fibroblast(F6) and UVB light(30 mJ/$cm^2$) was used to damage to dermal fibroblast. We measured the nitrite production, LDH release, and gene expression in UVB-irradiated dermal fibroblast to elucidate the actionmechanism of DS. Also, we evaluated the amount of increased PICP, TIMP-1 in dermal fibroblast. PICP, TIMP-1 concentration was measured using EIA kit, and gene expression (MMP-1, procollagen, c-fos, c-jun, NF-kB, Bcl-2, Bcl-xL, iNOS) were determined using real-time PCR. Results: 1. DS inhibited LDH-release, nitrite production in UVB-irradiated dermal fibroblast. 2. DS suppressed the gene expression of MMP-1 in UVB-irradiated dermal fibroblast. 3. DS increased the gene expression of procollagen in UVB-iradiated dermal fibroblast. 4. DS suppressed the gene expression of c-jun, c-fos, NF-kB, iNOS in UVBirradiated dermal fibroblast. 5. DS increased the gene expression of Bcl-2 in UVB-iradiated dermal fibroblast. 6. DS increased the cell proliferation of dermal fibroblast. Conclusions: From the results, we concluded DS increases the cell proliferation and collagen synthesis in dermal fibroblast. So we suggest that DS has the antiwrinkle effects.

• Biotechnology and Bioprocess Engineering:BBE
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• 제3권2호
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• pp.51-54
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• 1998

It has been proved that co-cultivation of human neroblastoma cells and human fibroblast cells can enhance nerve cell growth and the production of BDNF in perfusion cultivation. In batch co-cultivation, maximum cell density was increased up to 1.76${\times}$106 viable cells/mL from 9${\times}$105 viable cells/mL of only neuroblastoma cell culture. The growth of neuroblastoma cells was greatly improved by culturing both nerve and fibroblast cells in a perfusion process, maintaining 1.5${\times}$106 viable cells/mL, which was much higher than that form fed-batch cultivation. The nerve cell growth was greatly enhance in both fed-batch and perfusion cultivations while the growth of fibroblast cells was not. It strongly implies that the factors secreted from human fibrobast cells and/or the environments of co-culture system can enhance both cell growth and BDNF secretion. Specific BDNF production rate was not enhanced in co-cultures; however, the production period was increased as the cell growth was lengthened in the co-culture case. Competitive growth between nerve cells and fibroblast cells was not observed in all cases, showing no changes of fibroblast cell growth and only enhancement of the neuroblastoma cell growth and overall BDNF production. It was also found that the perfusion cultivation was the most appropriate process for cultivating two cell lines simultaneously in a bioreactor.

• 한국식품영양과학회지
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• 제26권6호
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• pp.998-1005
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• 1997

Kimchi showed protective effect from oxidative damage generated by hydrogen peroxide and paraquat. To investigate the major components of kimchi which reduce the cytotoxicity against reactive oxygen species, keratinocyte(A431, epidermoid carcinoma, human) and fibroblast(CCD-986SK, normal control, human) were cultured under oxidative stress condition provoked by paraquat, a superoxide anion generator, and hydrogen peroxide in the absence or presence of kimchi ingredients. Most keratinocyte and fibroblast cells were killed by hydrogen peroxide and paraquat over 1mM concentration, but kimchi ingredients showed protective effects from oxidative damage generated by hydrogen peroxide and onion, among those, garlic showed the most remarkable preventive effect. Most of kimchi ingredients showed protective effect against paraquat, especially leek notably increased cell survival. For fibroblast cells, ginger had the preventive effect against paraquat, especially leek notably increased cell survival. For fibroblast cells, ginger had the preventive effect from cell killing by high dose of hydrogen peroxide, but most ingredients were not effective against paraquat.

• Journal of Periodontal and Implant Science
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• 제36권3호
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• pp.783-796
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• 2006

The periodontium is a topographically complex organ consisting of epithelial tissue, soft and mineralized tissues. Structures comprising the periodontium include the gingiva, periodontal ligament (PDL) , cementum and the alveolar bone. The molecular mechanism of differentiation in PDL fibroblast cells remain unclear. Amino acid transporters play an important role in supplying nutrition to normal and cancer cells and for cell proliferation. Amino acid transport system L is a major nutrient transport system responsible for the Na+-independent transport of neutral amino acids including several essential amino acids. The system L is divided into two major subgroups, the L-type amino acid transporter 1 (LAT1) and the L-type amino acid transporter 2 (LAT2). In this study, the expression pattern of amino acid transport system L was, therefore, investigated in the differentiation of PDL fibroblast cells. To determine the expression level of amino acid transport system L participating in intracellular transport of amino acids in the differentiation of PDL fibroblast cells, it was examined by RT-PCR, observation of cell morphology, Alizaline red-S staining and uptake analysis after inducing experimental differentiation in PDL fibroblast cells isolated from mouse molar teeth. The results are as follows. 1. The LAT1 mRNA was expressed in the early stage of PDL fibroblast cell differentiation. This expression level was gradually reduced by differentiation- inducing time and it was not observed after the late stage. 2. The expression level of LAT2 mRNA was increased in time-dependent manner during differentiation induction of PDL fibroblast cells. 3. There was no changes in. the expression level of 4F2hc mRNA, the cofactor of LAT1 and LAT2, during differentiation of PDL fibroblast cells. 4. The expression level of ALP mRNA was gradually increased and the expression level of Col I mRNA was decreased during differentiation of PDL fibroblast cells. 5. The L-leucine transport was reduced by time from the early stage to the late stage in PDL fibroblast cell differentiation. As the results, it is considered that among neutral ammo acid transport system L in differentiation of PDL fibroblast cells, the LATl has a key role in cell proliferation in the early stage of cell differentiation and the LAT2 has an important role in the late stage of cell differentiation for providing cells with neutral amino acids including several essential amino acids.

• 대한한방부인과학회지
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• 제24권4호
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• pp.10-19
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• 2011

Objectives: This study was performed to elucidate the effects of Kwibi-tang extract(KB) on dermal fibroblast. Methods: To demonstrate the effects of KB on dermal fibroblast, we used human dermal fibroblast(F6) and UVB light(30 $mJ/cm^2$) was used to damage to dermal fibroblast. we measured the nitrite production, LDH release in UVB-irradiated dermal fibroblast to elucidate the action-mechanism of KB. Also, we evaluated cell proliferation of dermal fibroblast and the amount of increased PICP, TIMP-1 in dermal fibroblast. Results: 1. KB decreased the cell proliferation of F6 dermal fibroblast in concentration of 50 ${\mu}g/ml$. 2. KB decreased the synthesis of PICP in concentration of 50 ${\mu}g/ml$. 3. KB decreased the synthesis of TIMP-1 in concentration of 50 ${\mu}g/ml$. 4. KB have no effect on the damage in UVB-irradiated F6 dermal fibroblast. Conclusions: From the results, we concluded KB decreases the cell proliferation and collagen synthesis in dermal fibroblast. So we suggest that KB has the anti-hyperplasy of dermal fibroblast.

• Applied Microscopy
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• 제34권2호
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• pp.121-130
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• 2004

동물의 결합조직에 분포하고 있는 섬유모세포 (fibroblast)는 결합조직을 구성하는 세포의 한 종류로서 세포질 돌기들이 잘 발달된 형태적 특징이 있는 것으로 실험쥐 담관의 경우 간흡충 등의 기생충에 의하여 물리, 화학적 상해를 받았을 때 세포변이가 유발될 뿐만 아니라, 담관 암세포로 전이되기도 하는 것으로 알려져 있다. 따라서 저자 등은 실험쥐의 담관이 기생충에 의한 상해를 받았을 때 섬유모세포의 세포 표면과 세포질의 변화를 알아보고자 실험쥐 담관에서 섬유모세포를 분리하여 전자현미경으로 확인하고 다음과 같은 결과를 얻었다. 대조군 실험쥐 담관의 섬유모세포들은 일반적인 형태로 세포돌기, 세포표면 및 세포질을 구성하고 있었으나 간흡충 감염군 실험쥐 담관의 섬유모세포는 미세소관에 의한 세포질 돌기들이 다수 발달하고 다양한 종류의 포낭형 조면소포체 그리고 세포질에 전자밀도가 높은 다양한 액포, 높은 밀도의 리보좀을 포함하는 조면소포체, 다양한 형태의 과립 및 많은 수의 미세섬유가 관찰되는 형태적 변화가 관찰되었다. 간흡충에 감염된 담관의 섬유모세포는 간흡충에 의하여 상해 받은 세포가 물리화학적 자극에 의한 적응으로 단백질 합성이 증가하며 multi-vesicular 형태의 Golgi복합체가 생성되고, 세포질돌기 형성하는 것으로 확인되었다. 세포질에 광범위하게 분포하는 multi-vesicle은 당말단인 sialic acid를 포함하고 세포내에서 세포표면의 미세융모에 이르기까지 이동하는 것으로 확인되었다. 이상의 결과로 간흡충 감염 실험쥐로부터 분리된 섬유모세포는 actin단백으로 구성된 세포돌기가 잘 발달하고, 세포내 조면소포체에서 형성된 단백질이 Golgi복합체에서 당말단인 sialic acid로 전환되어 세포표면에 분포하게 된다. 이는 간흡충 감염으로 물리 화학적 자극 자극받은 섬유세포가 미세구조적 변화를 유발하는 것으로 확인되었다.

• Archives of Plastic Surgery
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• 제37권1호
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• pp.7-11
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• 2010

Purpose: Vascular endothelial growth factor (VEGF) is known as a growth factor of endothelium and fibroblast. The purpose is to know the VEGF effects on fibroblast proliferation and fibroblast's notch receptor expression. Methods: CCD-986sk fibroblast was purchased from the Korean Cell Bank and was used in XTT assay for proliferation and wound healing assay for migration. Immunofluorescent (IF) staining and western blotting were used in testing notch expression of fibroblast. Semiquantitative RT-PCR was used in checking notch 1 mRNA production by fibroblast. Student-t test was used for analyzing results. Results: Cell proliferation assay using XTT showed significant higher proliferation in VEGF treated fibroblast, $2.324{\pm}0.0026$ vs. $2.463{\pm}0.017$ (p=0.002). Wound healing assay showed longer migration in VEGF treated fibroblast (p=0.062). The fluorescence was brighter in VEGF treated cells of notch 1 IF staining. Notch 1 expressions and mRNA productions increased more in VEGF treated cells. Conclusion: VEGF stimulates fibroblast to proliferate, migrate and to express Notch 1 simultaneously. Notch receptor could be related to VEGF mediated wound healing.

• 대한한방부인과학회지
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• 제25권1호
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• pp.20-33
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• 2012

Objectives: This study was performed to elucidate the effects of Kanghwalsokdan-tang extract(KS) on hyper-plasy of collagen and cell damage in UVB-irradiated dermal fibroblast. Methods: To demonstrate the effects of KS on wound healing we used human dermal fibroblast(F6). We evaluated the amount of increased PICP, TIMP-1 in dermal fibroblast. PICP, TIMP-1 concentration was measured using EIA kit. Also, we measured the nitrite production, and LDH release in UVB-irradiated dermal fibroblast to elucidate the action-mechanism of KS. Results: 1. KS decreased the cell proliferation of dermal fibroblast. 2. KS decreased the biosynthesis of collagen in dermal fibroblast. 3. KS decreased the synthesis of TIMP-1 in dermal fibroblast. 4. KS had no effect on the LDH-release of UVB-irradiated dermal fibroblast. 5. KS inhibited nitrite production in UVB-irradiated dermal fibroblast. Conclusions: From the results, we concluded that KS has a protective effect on wound healing and photoaging.