• 한국미생물·생명공학회지
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• 제21권2호
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• pp.176-180
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• 1993

Survival and heat resistance of Listeria monocytogenes Scott A in various nutritional environments and cell type on stainless steel were determined. Viable cell of L. monocytogenes Scott A was most rapidly decreased in phosphate buffer among various media such as NSM (30 g TSB/1 l D.W.), LNM (2 g TSB/1 l D.W.) and phosphate buffer (pH=7) during incubation at 21 and 35C but survived for 15 days at 21C. Vegetative and starved L. monocytogenes Scott A were survived after heat treatment for 5 min at 65C while not detected at 72C.

• Asian Pacific Journal of Cancer Prevention
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• 제15권20호
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• pp.8947-8950
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• 2014

Immunological functions of heat shock proteins (HSPs) have long been recognized. In this study we aimed to efficiently purify HSP70 from renal cell carcinoma and test it as a tumor antigen for pulsing dendritic cells in vitro. HSP70 was purified from renal cell carcinoma specimens by serial column chromatography on Con A-sepharose, PD-10, ADP-agarose and DEAE-cellulose, and finally subjected to fast protein liquid chromatography (FPLC). Dendritic cells derived from the adherent fraction of peripheral blood mononuclear cells were cultured in the presence of IL-4 and GM-CSF and exposed to tumor HSP70. After 24 hours, dendritic cells were phenotypically characterized by flow cytometry. T cells obtained from the non-adherent fraction of peripheral blood mononuclear cells were then co-cultured with HSP70-pulsed dendritic cells and after 3 days T cell cytotoxicity towards primary cultured renal cell carcinoma cells was examined by Cell Counting Kit-8 assay. Dendritic cells pulsed in vitro with tumor-derived HSP70 expressed higher levels of CD83, CD80, CD86 and HLA-DR maturation markers than those pulsed with tumor cell lysate and comparable to that of dendritic cells pulsed with tumor cell lysate plus TNF-${\alpha}$. Concomitantly, cytotoxic T-lymphocytes induced by HSP70-pulsed dendritic cells presented the highest cytotoxic activity. There were no significant differences when using homologous or autologous HSP70 as the tumor antigen. HSP70 can be efficiently purified by chromatography and induces in vitro dendritic cell maturation in the absence of TNF-${\alpha}$. Conspecific HSP70 may effectively be used as a tumor antigen to pulse dendritic cells in vitro.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.115-118
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• 2000

미세 가공 기술을 이용하여 제작된 IDA 전극을 활용하여 임피던스 측정방식의 cell chip으로의 응용에 대해 고찰하였다. IDA 전극은 기존의 반도체 공정으로서 손쉽게 제작할 수 있고, 대량 제작시 전극의 재현성 확보가 용이하고 소형화 할 수 있으며 낮은 단가로 제작될 수 있는 장점이 있다. IDA 전극을 채용한 cell chip을 B16-F1 melanoma 세포 배양에 적용한 결과, 세포성장과 임피던스 변화량이 상관성을 보였고, 세포의 성장을 저해하는 약물의 투과시 cell chip의 임피던스 변화 역시 기존의 방법과 유사한 결과를 보여 주었다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권8호
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• pp.3397-3401
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• 2014

Determining cell quantity is a common problem in cytology research and anti-tumor drug development. A simple and low-cost method was developed to determine monolayer and adherent-growth cell quantities. The cell nucleus is located in the cytoplasm, and is independent. Thus, the nucleus cannot make contact even if the cell density is heavy. This phenomenon is the foundation of accurate cell-nucleus recognition. The cell nucleus is easily recognizable in images after fluorescent staining because it is independent. A one-to-one relationship exists between the nucleus and the cell; therefore, this method can be used to determine the quantity of proliferating cells. Results indicated that the activity of the histone deacetylase inhibitor Z1 was effective after this method was used. The nude-mouse xenograft model also revealed the potent anti-tumor activity of Z1. This research presents a new anti-tumor-drug evaluation method.

• 한국동물생명공학회지
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• 제38권3호
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• pp.121-130
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• 2023

Background: Coating a culture plate with molecules that aid in cell adhesion is a technique widely used to produce animal cell cultures. Extracellular matrix (ECM) is known for its efficiency in promoting adhesion, survival, and proliferation of adherent cells. Gelatin, a cost-effective type of ECM, is widely used in animal cell cultures including feeder-free embryonic stem (ES) cells. However, the optimal concentration of gelatin is a point of debate among researchers, with no studies having established the optimal gelatin concentration. Methods: In this study, we coated plastic plates with gelatin in a concentration-dependent manner and assessed Young's modulus using atomic force microscopy (AFM) to investigate the microstructure of the surface of each plastic plate. The adhesion, proliferation, and differentiation of the ESCs were compared and analyzed revealing differences in surface microstructure dependent on coating concentration. Results: According to AFM analysis, there was a clear difference in the microstructure of the surface according to the presence or absence of the gelatin coating, and it was confirmed that there was no difference at a concentration of 0.5% or more. ES cell also confirmed the difference in cell adhesion, proliferation, and differentiation according to the presence or absence of gelatin coating, and also it showed no difference over the concentration of 0.5%. Conclusions: The optimum gelatin-coating for the maintenance and differentiation of ES cells is 0.5%, and the gelatin concentration-mediated microenvironment and ES cell signaling are closely correlated.

• The Journal of Advanced Prosthodontics
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• 제8권3호
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• pp.235-240
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• 2016

PURPOSE. In this study, the aim of this study was to evaluate the effect of implant surface treatment on cell differentiation of osteoblast cells. For this purpose, three surfaces were compared: (1) a modified SLA (MSLA: sand-blasted with large grit, acid-etched, and immersed in 0.9% NaCl), (2) a laser treatment (LT: laser treatment) titanium surface and (3) a laser and acid-treated (LAT: laser treatment, acid-etched) titanium surface. MATERIALS AND METHODS. The MSLA surfaces were considered as the control group, and LT and LAT surfaces as test groups. Alkaline phosphatase expression (ALP) was used to quantify osteoblastic differentiation of MC3T3-E1 cell. Surface roughness was evaluated by a contact profilometer (URFPAK-SV; Mitutoyo, Kawasaki, Japan) and characterized by two parameters: mean roughness (Ra) and maximum peak-to-valley height (Rt). RESULTS. Scanning electron microscope revealed that MSLA (control group) surface was not as rough as LT, LAT surface (test groups). Alkaline phosphatase expression, the measure of osteoblastic differentiation, and total ALP expression by surface-adherent cells were found to be highest at 21 days for all three surfaces tested (P<.05). Furthermore, ALP expression levels of MSLA and LAT surfaces were significantly higher than expression levels of LT surface-adherent cells at 7, 14, and 21 days, respectively (P<.05). However, ALP expression levels between MSLA and LAT surface were equal at 7, 14, and 21 days (P>.05). CONCLUSION. This study suggested that MSLA and LAT surfaces exhibited more favorable environment for osteoblast differentiation when compared with LT surface, the results that are important for implant surface modification studies.

• IMMUNE NETWORK
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• 제7권3호
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• pp.117-123
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• 2007

Background: This study was designed to investigate the role of the hepatocyte growth factor (HGF) with regards to differentiation of somatic stem cells originating from the human umbilical cord blood (UCB) into hepatic lineage cells in vitro culture system. Methods: Mononuclear cells from UCB were cultured with and without HGF based on the fibroblast growth factor (FGF)-1, FGF-2, and stem cell factor. The cultured cells were confirmed by immunofluorescent staining analysis with albumin (ALB), cytokeratin-19 (CK-19), and proliferating cell nuclear antigen (PCNA) MoAb. ALB and CK-18 mRNA were also evaluated by reverse transcription-polymerase chain reaction. In order to observe changes in proliferating capacity with respect to the cultured period, CFSE with affinity to proliferating cells were tagged and later underwent flow cytometry. Results: In the HGF-treated group, cultured cells had a large oval shaped appearance with adherent, but easily detachable characteristics. In the HGF-non treated group, these cells were spindle-shaped with strong adherent characteristics. Expressions of ALB and CK-19 were evident in HGF-treated group compared to non-expression of those in to HGF-non treated group. Dual immunostaining analysis of the ALB producing cells showed presence of PCNA in their nuclei, and ALB and CK-18 mRNA were detected on the 21st day of cultured cells in the HGF-treated group. Conclusion: Our findings suggest that HGF has a pivotal role in differentiating somatic stem cells of human UCB into hepatic lineage cells in vitro.

• 한국진공학회지
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• 제7권s1호
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• pp.50-56
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• 1998

Ion beam assisted deposition (IBAD) was successfully used to produce a dense and ultra-adherent Hydroxyapatite (HA) film on titanium alloy and alumina. Recently it is also proved that the HA coatings on alumina substrate treated with 20 ppm $AgNO_3$ had the structure of $(Ag, Ca)_10(PO_4)6(OH)_2$, which exhibited excellent antimicrobial effects. The present paper aims to morphlogically characterize the adhesion of macrophages on newly developed Ag-HA coated alumina and Ti6A14V substrates and to evaluate the biocompatibility of the coatings in vitro. It can be found that the cell number on alumina of the concentration of $AgNO_3$ in the treatment, the cell number on Ag-HA coatings decreased. Up to 20 ppm $AgNO_3$ by Ag-treatment, the morphological development of the cells on Ag-HA coating was similar to that of the cells on HA coating, suggesting the biotolerance of the Ag-HA coatings.

• 한국표면공학회:학술대회논문집
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• 한국표면공학회 2013년도 춘계학술대회 논문집
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• pp.38-38
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• 2013

The mechanical cues that adherent cells derive from the extracellular matrix (ECM) can effect dramatic changes in cell migration, proliferation, and differentiation. Using a thin film of Type I collagen fibrils comprised of 100 nm to 200 nm collagen fibrils overlaying a bed of smaller fibrils, changes in cellular response to systematically controlled changes in mechanical properties of collagen was investigated. Further, an experimental and modeling approaches to calculate the elastic modulus of individual collagen fibrils, and thereby the effective stiffness of the entire collagen thin film matrix, from atomic force microscopy force spectroscopy data was performed. These results demonstrate an approach to analysis of fundamental properties of thin, heterogeneous, organic films, and add further insights into the mechanical properties of collagen fibrils that are of relevance to cell response to the ECM.

• 한국동물위생학회지
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• 제35권2호
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• pp.105-109
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• 2012

Apoptosis is a host defense mechanism that the cell uses to limit production of infectious pathogens. Although many bacteria, viruses and parasites can induce apoptosis in infected cells, some pathogens usually exhibit the ability to suppress the induction of apoptosis in the infected cells. Sophisticated evasion strategies of obligate intracellular parasites, in particular prevention of host cell apoptosis, are necessary to ensure successful replication. To study the ability of Eimeria tenella in this regard, in vitro experiments were performed applying Madin-Darby bovine kidney (MDBK) cells as host cell. We have demonstrated that productive infection of adherent cell lines by E. tenella resulted in an anti-apototic effect. This phenomenon was confirmed using in situ terminal deoxynucleotidyl transferase-mediated (TdT) deoxyuridine triphosphates (dUTP)-fluorescein nick end labeling (TUNEL) assay to detect apoptosis. Therefore, E. tenella could complete its cycle of productive infection while inducing anti-apoptosis in the infected cells. This finding might have implications for the pathobiology of E. tenella and other Eimeria species.