• Macromolecular Research
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• 제15권4호
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• pp.348-356
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• 2007

The aim of this study was to fabricate a submicro-and micro-patterned fibronectin coated wafer for a cell culture, which allows the positions and dimensions of the attached cells to be controlled. A replica molding was made into silicon via a photomask in quartz, using E-beam lithography, and then fabricated a polydimethylsiloxane stamp using the designed silicon mold. Hexadecanethiol $[HS(CH_2){_{15}}CH_3]$, adsorbed on the raised plateau of the surface of polydimethylsiloxane stamp, was contact-printed to form self-assembled monolayers (SAMs) of hexadecanethiolate on the surface of an Au-coated glass wafer. In order to form another SAM for control of the surface wafer properties, a hydrophilic hexa (ethylene glycol) terminated alkanethiol $[HS(CH_2){_{11}}(OCH_2CH_2){_6}OH]$ was also synthesized. The structural changes were confirmed using UV and $^1H-NMR$ spectroscopies. A SAM terminated in the hexa(ethylene glycol) groups was subsequently formed on the bare gold remaining on the surface of the Aucoated glass wafer. In order to aid the attachment of cells, fibronectin was adsorbed onto the resulting wafer, with the pattern formed on the gold-coated wafer confirmed using immunofluorescence staining against fibronectin. Fibronectin was adsorbed only onto the SAMs terminated in the methyl groups of the substrate. The hexa (ethylene glycol)-terminated regions resisted the adsorption of protein. Human dermal fibroblasts (P=4), obtained from newborn foreskin, only attached to the fibronectin-coated, methyl-terminated hydrophobic regions of the patterned SAMs. N-HDFs were more actively adhered, and spread in a pattern spacing below $14{\mu}m$, rather than above $17{\mu}m$, could easily migrate on the substrate containing spacing of $10{\mu}m$ or less between the strip lines.

• Advances in Traditional Medicine
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• 제2권2호
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• pp.119-124
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• 2002

Cigarette smoking contributes to lung cancer, cardiovascular diseases, oral diseases, etc. In desire to reduce their risk of disease, many cigarette smokers have tried to quit smoking. Sensory aspects of cigarette smoke are important for providing smoking satisfaction. Previously it was reported that citric acid aerosol significantly reduced craving for cigarettes and enhances smoking reduction and cessation. In this study, we tested whether a newly combined product Ciga-X, an aerosol for cessation aid, had toxicity in human embryonic lung fibroblast (MRC-9). The inhibitory effect of Ciga-X on cytotoxicity induced by cigarette smoke extract (CSE) or nicotine was examined in MRC-9, and craving for cigarettes and smorkers satisfaction after using Ciga-X was estimated. Ciga-X did not affect cell viability and had no toxicity in MRC-9. Ciga-X significantly inhibited not only CSE-induced cytotoxicity but also nicotine-induced cytotoxicity in MRC-9. One hundred and forty smokers rated the satisfaction for Ciga-X aerosol and craving reduction for cigarettes after using Ciga-X. The percentage of over 5 rating was 71.0% and 50.0% of subjects in satisfaction test for Ciga-X compared to their own brand and in craving reduction for cigarette, respectively. Besides, craving reduction for cigarette was highly correlated with the duration of smoking. Subjects have smoked under 10 years were more reduced in craving for cigarettes after using Ciga-X as compared to over 10 years (p=0.049). These results suggest that Ciga-X may be effective in promoting smoking abstinence with the reduction of CSE- or nicotine-induced human lung fibroblasts cytotoxicity.

• 대한의생명과학회지
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• 제23권3호
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• pp.238-250
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• 2017

Patient's primary tumor-derived tumor cell lines likely represent ideal tools for human tumor biology in vitro and in vivo. Here, we describe eight human gastric carcinoma cell lines derived from established tumors in vivo upon subcutaneous transplantation of primary gastric carcinoma specimens in BALB/c nude mice. These xenografted gastric tumor cell lines (GTX) displayed close similarity with primary gastric tumor tissues in their in vivo growth pattern and genomic alterations. GTX-085 cells were resistant to cisplatin, while GTX-087 was the most sensitive cell line. GTX-085 was the only cell line showing a metastatic potential. Epithelial cell adhesion molecule (EPCAM) expression was especially strong in all tissue samples, as well as in cell cultures. GTX-139, the largest tumor graft obtained after injection, displayed distinct expression of CD44v6, fibroblast growth factor receptor 2 (FGFR2), and prominin 1 (PROM1, also known as CD133). In summary, we established eight xenograft gastric cancer cell lines from gastric cancer patient tissues, with their histological and molecular features consistent with those of the primary tumors. The established GTX cell lines will enable future studies of their responses to various treatments for gastric cancer.

• Biomolecules & Therapeutics
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• 제24권6호
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• pp.572-580
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• 2016

3-Deoxysappanchalcone (3-DSC) has been reported to possess anti-allergic, antiviral, anti-inflammatory and antioxidant activities. In the present study, we investigated the effects of 3-DSC on the proliferation of human hair follicle dermal papilla cells (HDPCs) and mouse hair growth in vivo. A real-time cell analyzer system, luciferase assay, Western blot and real-time polymerase chain reaction (PCR) were employed to measure the biochemical changes occurring in HDPCs in response to 3-DSC treatment. The effect of 3-DSC on hair growth in C57BL/6 mice was also examined. 3-DSC promoted the proliferation of HDPCs, similar to Tofacitinib, an inhibitor of janus-activated kinase (JAK). 3-DSC promoted phosphorylation of ${\beta}$-catenin and transcriptional activation of the T-cell factor. In addition, 3-DSC potentiated interleukin-6 (IL-6)-induced phosphorylation and subsequent transactivation of signal transducer and activator of transcription-3 (STAT3), thereby increasing the expression of cyclin-dependent kinase-4 (Cdk4), fibroblast growth factor (FGF) and vascular endothelial growth factor (VEGF). On the contrary, 3-DSC attenuated STAT6 mRNA expression and IL4-induced STAT6 phosphorylation in HDPCs. Finally, we observed that topical application of 3-DSC promoted the anagen phase of hair growth in C57BL/6 mice. 3-DSC stimulates hair growth possibly by inducing proliferation of follicular dermal papilla cells via modulation of $WNT/{\beta}$-catenin and STAT signaling.

• Biomolecules & Therapeutics
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• 제18권1호
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• pp.56-64
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• 2010

Pentoxifylline (PTX) has been used for the local reduction of fat tissue in the clinical setting. However, its safety and efficacy have not been proven. The aim of this study was to evaluate the effects of PTX on cell lines established from fat tissue. Newly cultured human preadipocytes and adipocytes from subcutaneous abdominal fat in addition to purchased human lung fibroblasts and keratinocytes were treated with PTX at different concentrations. Cell viability was determined using the Cell counting kit (CCK)-8 assay and lipolysis was evaluated using an Elisa kit. DNA fragmentation, Western blot analysis, Hoechst and Propidium Iodide (PI) staining and fluorescence activated cell scanning analysis were performed to confirm apoptosis. The viability of adipocytes, preadipocytes, keratinocytes and fibroblasts was markedly decreased at concentrations of PTX above 20 mM. Apoptosis was induced at concentrations of PTX over 40 mM in all cell lines. Lipolysis was increased by 60% at concentrations of PTX of 20 mM compared to the control. In conclusion, the results of this study showed that 20 mM of PTX induced lipolysis. At concentrations over 20 mM, PTX reduced the viability of all cells studied including: adipocytes, preadipocytes, fibroblasts and keratinocytes, in a non-specific manner.

• Asian Pacific Journal of Cancer Prevention
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• 제17권7호
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• pp.3281-3287
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• 2016

Bioactive compounds extracted from leaves and twigs of Goniothalamus griffithii include pinocembrin (PCN) and goniothalamin (GTN). The objectives of this study were to investigate the cytotoxic activities of PCN and GTN and their influence on molecular signaling for cell death in several human cancer cell lines compared to normal murine fibroblast NIH3T3 cells. GTN exhibited the most potent cytotoxicity against MCF-7 > HeLa > HepG2 > NIH3T3 cells with $IC_{50}$ values of 7.33, 14.8, 37.1 and $65.4{\mu}M$, respectively, whereas PCN was cytotoxic only to HepG2 cells with $IC_{50}$ values of ${\sim}80{\mu}M$. Apoptotic cell death was confirmed by staining the cells with annexin V-FITC and propidium iodide (PI) employing flow cytometry. Apoptosis was shown by externalization of phosphatidylserine in goniothalamin-treated MCF-7 cells in a dose response manner. Positive PI-stained cells with the typical morphology of apoptotic cells were increased dose-dependently. Furthermore, reduction of mitochondrial transmembrane potential was found in goniothalamin-treated MCF-7, HepG2 and HeLa cells. GTN treatment in MCF-7 increased caspase-3, -8 and -9 activities while GTN-induced HeLa cells showed an increase of both caspase-3 and -9 activities. But an increased caspase-8 activity was demonstrated in GTN- and PCN-treated MCF-7 and HepG2 cells, respectively. Taken together, GTN- and PCN-induced human cancer cell apoptosis was through different molecular mechanisms or signaling pathways, which might be due to different machineries in different types of cancer cells, as evidenced by the compound-modulated caspase activities in both intrinsic and/or extrinsic pathways.

• 한국가금학회:학술대회논문집
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• 한국가금학회 2001년도 제18차 정기총회 및 학술발표 PROCEEDINGS
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• pp.40-46
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• 2001

The use of pluripotent stem cells has tremendous advantages for various purposes but these cell lines with proven germ-line transmission have been completely established only in the mouse. Embryonic germ (EG) cell lines are also pluripotent and undifferentiated stem cells established from primordial germ cells (PGCs). This study was conducted to establish and characterize the chicken EG cells derived from gonadal primordial germ cells. We isolated gonadal PGCs from 5.5-day-old (stage 28) White leghorn (WL) embryos and established chicken EG cells lines with EG culture medium supplemented with human stem cell factor (hSCF), murine leukemia inhibitory factor (mLIF), bovine basic fibroblast growth factor (bFGF), human interleukin-11 (hIL-11), and human insulin-like growth factor-I (hIGF-I). These cells grew continuously for 4 months (10 passages) on a feeder layer of mitotically active chicken embryonic fibroblasts. These cells were characterized by screening with the Periodic acid-Shiff＇s reaction, anti-SSEA-1 antibody, and a proliferation assay after several passages. As the results, the chicken EG cells maintained characteristics of undifferentiated stem cells as well as that of gonadal PGCs. When cultured in suspension, the chicken EG cells successfully formed an embryoid body and differentiated into a variety of cell types when re-seeded onto culture dish. The chicken EG cells were injected into blastodermal layer at stage X and dorsal aorta of recipient embryo at stage 14 (incubation of 53hrs) and produced chimeric chickens with various differentiated tissues derived from the EG cells. The germline chimeras were also successfully induced by using EG cells. Thus, Chicken EG cells will be useful for the production of transgenic chickena and for studies of germ cell differentiation and genomic imprinting.

• 한국축산식품학회지
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• 제23권2호
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• pp.155-160
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• 2003

인유 초유로부터 지방을 제거하여 skim milk을 얻었다. 이것을 ultrafiltration을 한 후 gel filtration을 거쳐 EGF fraction 을 획득하였으며, 이를 SDS-PAGE로 확인하였다. hish performance liquid chromatography(HPLC)에 의해 EGF fraction을 분석한 결과, standard EGF 분석과 비교했을 때 31.545분대와 유사한 31.185분대에 peak가 나타난 것을 인하였다. 분리한 fraction 중 EGF 함유량을 측정하기 위해 immunoassay를 실시한 결과 건조중량 10 mg/mL 중에 약 10ng의 EGF가 포함되어 있는 것으로 측정되었다. SDS- PAGE후 실시한 western blot을 통하여 분리한 분획의 분리도를 확인하였다. 세포성장에 미치는 영향을 살펴본 결과, fibroblast cell인 BALB/3T3의 경우에는 건조중량 1 mg/mL 농도의 fraction 처리후 약 95%의 성장률을 보였으며, epithelial cell 인 IEC-6의 경우에는 약 71%의 성장률을 보였다.

• Restorative Dentistry and Endodontics
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• 제8권1호
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• pp.7-17
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• 1982

The purpose of this study was to investigate the fine structural modifications of fibroblasts in the coronal region of inflamed human pulps from carious teeth. Six untreated human teeth with large carious lesions and two normal teeth as control were selected from male and female patients between the ages of 20 and 39. The teeth were divided into 4 groups by light microscopic findings: the normal control group, the chronic inflammatory cell-appeared group, the acute and chronic inflammatory cell-appeared group, and the total necrosis group. All tissues were fixed in 2.5% glutaraldehyde in 0.1M sodium cacodylate buffer at pH 7.4 and 1% osmic acid in same buffer. They were embedded in Epon 812. The ultrathin sections were stained conventionally and examined with a AEI Corynth 500 electron microscope. The results were as follows; 1. The fibroblasts of the normal pulps were almost in a quiescent state. 2. The active and the quiescent fibroblasts were found in the pulps of the chronic inflammatory cell-appeared group. Lymphocytes and plasma cells were also seen scattered among these fibroblasts. 3. In the pulps of the acute and chronic inflammatory cell-appeared group, active, degenerative and necrotic fibroblasts were found in the PMN appeared area. And all the fibroblasts in the fibrosis area were active. In the area of chronic inflammatory cellular infiltration, almost all the fibroblasts were active, but seldom were quiescent fibroblasts observed. Some fibroblasts in the pulps of two teeth had large vacuoles that contained banded collagen fibrils. The phagosomes had small beaded vesicles or large lysosome-like varicosity. In two of the teeth, microorganisms were present and two morphological shapes were identified, a rod and a coccus. 4. Vacuolar, vesicular, lamellar, fibrous and myelin structures were observed in the pulp of the total necrosis group, and cocci were also seen.

• Journal of Ginseng Research
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• 제44권5호
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• pp.738-746
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• 2020

Background: Red ginseng contains components, including microelements, vitamins, essential oils, and fatty acids, that can be used in skincare to delay the aging process. We investigated the effects of red ginseng treatment on skin elasticity by assessing cellular stiffness and measuring collagen protein synthesis. Methods: Human dermal fibroblasts were treated with red ginseng, and the resulting changes in stiffness were investigated using atomic force microscopy. Cytoskeletal changes and mRNA expression of biomarkers of aging, including that of procollagens I and VII, elastin, and fibrillin-1, were investigated. Collagen in a human skin equivalent treated with red ginseng was visualized via hematoxylin and eosin staining, scanning electron microscopy, and atomic force microscopy. Results and conclusion: The stiffness of fibroblasts was significantly reduced by treatment with red ginseng concentrations of ≥ 0.8 mg/mL. The ratio of F-actin to G-actin decreased after treatment, which corresponded to a change in fibroblast stiffness. The storage modulus (G') and loss modulus (G'') of the skin equivalent were both lowered by red ginseng treatment. This result indicates that the viscoelasticity of the skin equivalent can be restored by red ginseng treatment.