• Reproductive and Developmental Biology
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• 제39권4호
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• pp.127-132
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• 2015

To overcome the hyperacute immune rejection during pig-to-non-human primates xenotranasplantation, we have produced and bred ${\alpha}$-1,3-galactosyltransferase knock-out ($GalT^{-/-}$) pigs. In this study, the somatic cells and tissues from the $GalT^{-/-}$ pigs were characterized by an analysis of the expression of Gal${\alpha}$-1,3-Gal (${\alpha}-Gal$) epitope. Briefly, ear fibroblast cell lines of 19 homozygous $GalT^{-/-}$ pigs were established and cryopreserved. The expression of ${\alpha}-Gal$ epitope in the cells was measured by fluorescence activated cell sorter (FACS) analysis using BS-I-B4 lectin. Also, the homozygous ($GalT^{-/-}$) cells and tissues samples were immunostained with BS-I-B4 lectin for analysis of ${\alpha}-Gal$ epitope expression. The results showed that the expression of ${\alpha}-Gal$ epitope in $GalT^{-/-}$ cells (0.2 %) were significantly (p<0.05) down-regulated to the range of cynomolgus monkey fibroblast (0.2 %) cells compared to heterozygous ($GalT^{-/+}$) (9.3 %) and wild type ($GalT^{+/+}$) (93.7 %) fibroblast cells. In the immunostaining results, while the expression of ${\alpha}-Gal$ epitope was detected a partly in $GalT^{-/+}$ cells and mostly in $GalT^{+/+}$ cells, it was almost not detected in the $GalT^{-/-}$ cells. Also, immunostaining results from various tissues of the $GalT^{-/-}$ pig showed that the expression of ${\alpha}-Gal$ epitope was not detectable, whereas various tissues from $GalT^{+/+}$ pig showed a strong expression of ${\alpha}-Gal$ epitope. Our results demonstrated that ${\alpha}-Gal$ epitope expressions from $GalT^{-/-}$ pigs were successfully knocked out to prevent hyperacute immune rejection for further study of xenotransplantation.

• Bulletin of the Korean Chemical Society
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• 제32권9호
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• pp.3405-3410
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• 2011

We have investigated the cytotoxic potential of tellurium (Te) nanowires in BALB/3T3 fibroblast cells. Te nanowires were synthesized through an aqueous phase surfactant assisted method. Toxicological experiments, such as analysis of morphological changes, MTT assay, DAPI staining, and estimation of intracellular reactive oxygen species, were carried out to reveal the cytotoxic effects of Te nanowires. Te nanowires were found to be cytotoxic at all concentrations tested, in a dose-dependent manner. The UV/Vis spectra of Te nanowires suspended in a culture medium showed drastic changes and disappearance of two broad absorption peaks. The physicochemical properties such as, surface charge, size, and shape of Te nanowires were found to be altered during exposure of cells, due to the instability and agglomeration of nanowires in the culture medium. These results suggest that the chemical components of the DMEM medium significantly affect the stability of Te nanowires. In addition, TEM images revealed that necrosis was the basic pattern of cell death, which might stem from the formation of toxic moieties of tellurium, released from nanowire structures, in the bioenvironment. These observations thus suggest that Te nanomaterials may pose potential risks to environmental and human health.

• 한국환경농학회지
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• 제16권2호
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• pp.149-155
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• 1997

Carbofuran에 의하여 손상된 NIH3T3 섬유모세포(mouse skin fibroblast)를 보상시킬 수 있는 물질을 개발하기 위한 기초자료를 제시하고자 carbofuran이 NIH3T3 섬유모세포에 미치는 세포독성을 검정하고, carbofuran의 독성에 대한 phenobarbital sodium(PB) 및 3-methylcholantherene(3-MC)의 보상효과를 비교${\cdot}$분석하였다. Carbofuran의 $IC_{50}$ 결정은 배양중인 NIH3T3 섬유모세포의 각 well당 1, 25, 50, $100{\mu}M$의 carbofuran을 첨가하여 48시간 배양한 후 MTT(Tetrazolium MTT), NR(Neutral red) 및 SBR(Sulforhodamine B protein)정량을 실시하여 이들에 대한 각각의 $IC_{50}$을 구하였으며, 여기에서 구한 carbofuran의 $IC_{50}$농도와 여러 농도의 PB 또는 3-MC를 배양액에 첨가하여 48시간 배양한 후 MTT, NR 및 SRB 정량을 실시하여 보상효과를 측정하고 광학현미경적 관찰을 실시하였다. Carbofuran의 세포독성 실험결과를 보면 MTT흡광도는 carbofuran의 농도증가에 따라 감소하였으며 $MTT_{50}$은 $60.7{\mu}M$이었고, NR흡광도는 $100{\mu}M$농도에서 급격히 감소하였으며, $NR_{50}$은 $82.5{\mu}M$이었고, SRB흡광도는 $50{\mu}M$농도에서 급격히 감소하였으며 $SRB_{50}$은 $87.0{\mu}M$로써 50%의 세포독성을 나타냈다. 보상효과 실험에서는 carbofuran IC50과 PB의 조합처리의 경우 MTT 정랑과 NR정량에서는 유사하게 PB $100{\mu}M$처리군에서부터 유의성있는 보상효과가 나타났으나, SRB정량에서는 보상효과가 인정되지 않았다. Carbofura $IC_{50}$과 3-MC의 조합처리의 경우 MTT정량은 3-MC $50{\mu}M$처리군에서부터, NR정량과 SRB정량의 경우는 동일하게 3-MC $100{\mu}M$처리군에서부터 유의성있는 보상효과가 나타났다. 세포의 광학현미경적 관찰소견의 경우에서도 carbofuran과 PB 또는 3-MC 조합처리 실험군 모두에서 세포가 회복되는 것을 관찰할 수 있었다. 이상의 결과에서 PB와 3-MC 모두 carbofuran의 세포독성을 감소시킬 수 있는 물질임을 알 수 있었다.

• Archives of Pharmacal Research
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• 제19권1호
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• pp.36-40
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• 1996

The effect of glycyrrhetinic acid(18.betha.-glycyrrhetinic acid, GA) on histamine synthesis and release was investigated in cocultured mast cells with Swiss 3T3 fibroblasts. GA has strong dose dependent inhibitory activity for histamine synthesis and release in cocultured mast cells. GA(50 .mu.M) inhibited about 85% of histidine decarboxylase (HDC) activity. The appearance of cells staining positively with berberine sulface was also decreased in the presence of GA. It indicates that transdifferentiation of cultured mas cells (CMCs) was also inhibited.

• 한국지역사회생활과학회지
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• 제15권1호
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• pp.67-76
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• 2004

In order to promote the value of the flowers as new agricultural products, we investigated the biological activities of rape, arrowroot, and rose extracts. Biological activities investigated included antioxidant activity and the effects on 3T3-L1 fibroblast cells. When each flower was extracted with methanol, the antioxidant index and electron donating activity of roses was the highest $(IC_{50}$ of rose extract was $17.6 \mu{g}/m\ell$). When 3T3-L1 fibroblast cells were treated with extracts made with hexane, ethyl acetate, and ether, the rape extracts had a cytotoxic effect on the cells. 12.2% of cells survived when treated with a 3mg/$m\ell$ ether extract while those treated with the same concentration of hexane and ethyl acetate had survival rates of 76.2% and 78.6% respectively. In contrast to rape, the ether extract of arrowroot and rose stimulated the growth of 3T3-L1 cells. The effect of rose extracts was much bigger than those of other extracts. Although every rose extract stimulated the growth of the 3T3-L 1 cells, the ether extract stimulated growth up to 168.6% compared to the control at the concentration of $0.3mg/m\ell$, and 148.3% at the concentration of $1mg/m\ell$. The toxicity on cells treated with $H_2 O_2$ of $450\mu{M}l$was decreased with the addition of rose extract. The survival rate after treatment with rose extract at the concentration of $100\mu{g}/m\ell$ was increased to 71% compared to the 32% survival rate of control. From these results, it can be concluded that the extracts of arrowroot and rose seem to stimulate cells, whereas the extract of rape has a cytotoxic effect. Biological activities of ether extract were the strongest compared to those of other extracts at the tested concentrations.

• 대한약학회:학술대회논문집
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• 대한약학회 2001년도 Proceedings of the Pharmaceutical Society of Korea
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• pp.169.2-170
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• 2001

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2003년도 제39회 학술심포지움
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• pp.144-144
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• 2003

• BMB Reports
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• 제29권3호
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• pp.205-209
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• 1996

To investigate the role of heparin in keratinocyte growth factor (KGF) and acidic fibroblast growth factor (aFGF) high affinity binding to the KGF receptor (KGFR), a cell free system was established which utilized a secreted chimeric molecule between the KGFR extracellular domain and the immunoglobulin heavy chain Fc domain (KGFR-HFc). KGFR-HFc was purified from NIH 3T3 cells and demonstrated the binding of $[^3H]-heparin$ as well as heparin Sepharose. Scatchard analysis showed that the dissociation constant for heparin binding to KGFR-HFc was 140 nM. High affinity KGF and aFGF binding to KGFR-HFc remained unchanged after treatment with 0.6 M NaCl, which is the concentration sufficient to release any bound heparin to the KGFR-HFc. These results strongly suggest that although the KGFR interacts with heparin, the presence of heparin is not absolutely required for high affinity binding of either KGF or aFGF to the KGFR.

• KSBB Journal
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• 제11권2호
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• pp.254-261
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• 1996

수두 바이 러스(Varicella-zoster virus, VZV) 백신을 생산하기 위하여 약독화된 수두 바이러스 주인 Oka 주를 사용하여 정상세포인 인체 폐세 포(human lung fibroblast cells)로부터 수두 바이 러스 생산조건을 검토하였다. 숙주세포의 배양을 위 한 최적 성장배지로는 DMEM이 조사된 배지 중에 서는 가장 좋음을 알 수 있었으며 숙주세포의 성장율은 세포 age의 지표인 population doubling level (PDL) 이 46 이상인 경우 낮아지는 것무로 확인되 었으며 따라서 수두바이러스 생산을 위해서는 PDL 4 46 이하의 young cell을 사용해야 할 것으로 사료된 다 한편 수두바이러스 생산을 위한 최적 증식조건 으로는 배양온도 $37^{\circ}C$, 혈청농도 2%, MOl 1:5였다. 숙주세포를 대량배양하기 위한 기본조건을 검토 하기 위하여 미 럽담체 (Cytodex-3)를 이용한 spm­n ner flask culture를 시행하였다. 미럽담체에서의 숙 주세포 성장은 T -flask와 비하여 별다른 차이가 없었으나 VZV의 증식은 T-flask 배양에서보다 훨씬 낮았다. 이는 미립담체배양에서의 높은 전단응력 과 미립담체끼리의 영김현상 때문으로 생각된다.

• 대한화장품학회지
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• 제35권1호
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• pp.19-25
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• 2009

피부주름 개선에는 콜라겐(collagen)뿐만 아니라 탄력섬유인 엘라스틴(elastin) 등도 기여하는 것으로 보고되고 있다. 더 나아가 자외선에 의하여 사람의 피부에서 광노화 현상이 나타나며 자외선 조사 후 엘라스타제(elastase)의 활성이 증가하기 때문에 엘라스타제의 활성증가는 자외선에 의한 피부 탄성도의 감소 및 주름 생성의 주요원인으로 생각된다. 따라서 본 연구에서는 피부주름 생성에 영향을 미치는 엘라스타제의 활성을 측정하기 위한 모델을 마련하기 위하여 시판되는 두 가지 엘라스타제, 돼지 췌장 엘라스타제(porcine pancreatic elastase)와 사람 호중구 엘라스타제(human neutrophil elastase)를 사용하였으며 다음 세 가지는 정상 사람 섬유아세포(normal primary human fibroblasts), 쥐의 3T3 섬유아세포주(3T3 mouse fibroblasts), 사람의 CCD-25Sk 섬유아세포주(CCD-25Sk human fibroblasts)로부터 elastase를 준비하여 사용하였다. 준비된 5가지 효소의 농도에 기질의 농도 및 배양시간에 따르는 효소의 활성을 비교 평가하였다. 양성대조군으로 사용한 phosphoramidon이 normal human primary fibroblast elastase와 CCD-25Sk fibroblast elastase의 활성을 유의성 있게 억제한 반면에 porcine pancreatic elastase에는 별다른 영향을 미치지 못하였다. 따라서 주름 개선 후보물질 탐색을 위한 엘라스타제의 선정에는 신중한 고려가 필요할 것으로 사료된다.