• 대한세포병리학회:학술대회논문집
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• 1992.06a
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• pp.7.1-7.1
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• 1992

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2012.05a
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• pp.909-912
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• 2012

An automated cell tracking system is automatically to analyze and track changes of cell behaviors in time-lapse cell images acquired from microscope in the cell culture. In this paper, we proposed and developed an ellipse fitting based algorithm for separating very small size overlapping cells in a cell image consisted of thousands or ten thousands cells. We were extracted contours of clusters and divided them into line segments and then produced their fitted ellipses for each line segment. By experimentations, our algorithm was separated clusters with average 91% precision for two overlapping cells and average 84% precision for three overlapping cells respectively.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.73-73
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• 2003

본 연구는 돼지 체세포의 clonal cell lines을 효율적으로 확립할 수 있는 배양방법을 제시하기 위하여 실시하였다. 50일령의 돼지태아로부터 섬유아세포를 회수하여 2회 계대배양 후 단일 세포를 96-well plate와 4-well dish에서 배양하여 배양액내의 FBS농도(10, 30, 50%), 첨가제(catalase, $\beta$-mercaptoethanol; BME), 세포의 크기(<16, 16~20, 20<) 및 형태(smooth, rough)에 따른 cell line 확립 효율을 검토하였다. FBS 농도에 따른 clonal line 확립 효율과 PD를 검토한 결과, 효율에 있어서 30% FBS 처리구(5.1%)가 10%(0.3%) 및 50%처리구(2.1%)보다 비교적 높게 나타났으나 유의적 차이는 없었으며, PD의 경우는 10, 30, 50%처리구에서 각각 36.7, 29.4, 26.3 시간으로 FBS 농도가 증가할수록 PD 시간이 유의적으로 짧아졌다(p<0.05). 배양액 첨가제에 있어서 BME 가시 무처리구나 catalase 첨가시보다 유의적으로 높은 효율을 보였으며(11.4%, 3.2%, 3.2%; p<0.05), PD 시간은 짧게 나타났다(23.6, 25.5, 28.1; p<0.05). 그러나 catalase는 cell line 확립 효율에 영향을 미치지 않는 것으로 나타났다. 세포의 크기에 따른 cell line 확립효율은 5.2~8.2%로 크기에 따른 차이는 보이지 않았으며, PD 또한 23.7~27.9 시간으로 세포 크기에 따른 차이는 없었다. 세포의 형태에 따른 세포의 부착율은 smooth(55.8%)구가 rough(73.0%)구보다 유의적으로 낮게 나타났으나(p<0.05), clonal line 확립 효율(7.7%n vs. 6.6%) 및 PD(23.5h vs. 24.0h)에서는 차이가 없었다. 본 연구의 결과, 세포의 형태에 따른 cell line 확립 효율은 큰 차이가 없었으나, 세포 배양액내에 30% FBS와 BME의 첨가로 clonal cell line 확립 효율을 향상시킬 수 있음을 확인할 수 있었다. 또한 cell line 확립 효율이 높아질수록 PD 시간이 짧아지는 경향을 볼 수 있었다.

• Korean Journal of Animal Reproduction
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• v.19 no.4
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• pp.293-305
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• 1996

Retrovirus는 DNA가 아닌 RNA를 유전 물질로 갖고 있는 동물 virus인데 각 virus는 RNA와 함께 크게 gag, pol. 그리고 env 등의 3가지 단백질로 구성되어 있다. gag 단백질은 virus의 내부구조를 형성하는 단백질이고, pol단백질은 감염을 통해 표적 세포에 도입된 retrovirus의 RNA를 DNA로 역전사시키는 reverse transcriptase의 역할을 하며, env단백질은 virusdml 외부를 구성하는 단백질로써 이 단백질에 의해 각 retrovirus의 종류에 따른 감염이 가능한 표적세포의 종류가 결정된다(host cell specificity). 따라서 어떤 retrovirus의 envelope단백질과 표식세포에 있는 retrovirus의 envelope 단백질에 대한 특정 receptor와의 상호 작용에 의해 세포속으로 도입된 virus의 RNA는 reverse transcriptase에 의해 DNA로 역전사된 후 표적세포의 genomic DNA에 삽입되는 특징을 가진다. 이러한 특징을 가진 retrovirus vector system은 형질 전환 동물의 생산에 있어서 현재까지의 주된 방법인 수정란의 pronucleus에의 DNA microinjection방법 보다 여러 가지 면에서 우수함에도 불구하고 쥐 이외의 다른 동물에서는 거의 이용되고 있지 않는 실정이다. 주된 원인으로는 현재 사용되고 있는 대부분의 retrovirus vector system이 쥐의 백혈병 virus를 근간으로 하기 때문에 이 system에서 생산된 virus는 쥐 이외의 다른 동물, 특히 유제류의 세포에는감염성이 아주 약하기 때문이다. 이러한 결점을 해결하기 위하여 최근에 기존의 쥐 백혈병 virus의 envelope protein을 vesicular stomatitis virus의 G protein으로 대체한 hybrid retrovirus vector system이 개발되었다. 이러한 system에서 생산되는 virus는 조류를 포함한 거의 모든 종류의 동물세포를 감염시킬 수 있으며 몇몇 특정세포에 대해서는 기존의 retrovirus vector system에 비해 1,000배 이상의 높은 감염도를 나타내는데 그 특징이 있다. 따라서 이러한 새로운 virus vector system을 이용할 경우, 보다 다양한 종에 있어서 형질전환 동물을 효율적으로 생산할 수 있을 뿐만 아니라 형질전환 동물의 생산 방법 자체를 다양화 시킬 수 있다고 본다.

• Journal of Life Science
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• v.28 no.8
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• pp.992-998
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• 2018

Cancer cells grow in an environment composed of various components that supports tumor growth. Major cell types in the tumor microenvironment are fibroblast, endothelial cells and immune cells. All of these cells communicate with cancer cells. Among infiltrating immune cells as an abundant component of solid tumors, macrophages are a major component of the tumor microenvironment and orchestrates various aspects of immunity. The complex balance between pro-tumoral and anti-tumoral effects of immune cell infiltration can create a chronic inflammatory microenvironment essential for tumor growth and progression. Macrophages express different functional programs in response to microenvironmental signals, defined as M1 and M2 polarization. Tumor-associated macrophages (TAM) secret many cytokines, chemokines and proteases, which also promote tumor angiogenesis, growth, metastasis and immunosuppression. TAM have multifaceted roles in the development of many tumor types. TAM also interact with cancer stem cells. This interaction leads to tumorigenesis, metastasis, and drug resistance. TAM obtain various immunosuppressive functions to maintain the tumor microenvironment. TAM are characterized by their heterogeneity and plasticity, as they can be functionally reprogrammed to polarized phenotypes by exposure to cancer-related factors, stromal factors, infections, or even drug interventions. Because TAMs produce tumor-specific chemokines by the stimulation of stromal factors, chemokines might serve as biomarkers that reflect disease activity. The evidence has shown that cancer tissues with high infiltration of TAM are associated with poor patient prognosis and resistance to therapies. Targeting of TAM in tumors is considered a promising therapeutic strategy for anti-cancer treatment.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.45 no.4
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• pp.373-380
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• 2019

There are many differences in tran-epidermal water loss (TEWL), skin water contents, and skin elasticity, etc between face and forearm skin. In particular, our previous studies showed that elasticity of face skin was significantly differed from forearm depending on full hydration. So, we have studied the surface properties of corneocyte using atomic force microscopy (AFM), assuming that the differences between face and forearm skin would be associated with the surface properties of corneocyte. The surface roughness of corneocyte and villus-like projections (VPs) were measured. Furthermore, qualitative comparison among the surface of face, forearm, and lip corneocyte was performed. Corneocytes were collected by tape-stripping on both face and forearm of 8 volunteers, and the bottom surface of corneocytes were measured at 40 ㎛ × 40 ㎛ using AFM. Results showed that the lower surface roughness of face corneocytes was 388.34 ± 86.189 nm, and that of forearm corneocytes was 662.27 ± 224.257 nm, which confirmed that the lower surface of forearm corneocytes was more rough than that of face corneocytes (p < 0.001). Compared with the amount of VPs, lip corneocytes were the highest followed by face corneocytes, and forearm corneocytes were the lowest. From these results, it is conclued that the surface properties of corneocytes are somewhat involved in the property differences between the face and the forearm skin and VPs can be a useful parameter for the study of corneocyte by site. In addition, AFM is a very useful device for the comparative study of nano-structural differences on the surface of corneocytes. More studies can lead to develop a new evaluation method of corneocytes.

• Korean Chemical Engineering Research
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• v.46 no.6
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• pp.1100-1106
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• 2008

In this study, we presented rapid and simple fabrication method of functionalized surface on various substrates as a universal platform for the selective immobilization of cells. The functionalized surface was achieved by using deposition of polyelectrolyte such as poly(allyamine hydrochloride) (PAH), poly(diallyldimethyl ammonium chloride) (PDAC), poly(4-ammonium styrene sulfonic acid) (PSS), poly(acrylic acid) (PAA) and fabrication of poly(ethylene glycol) (PEG) microstructure through micro-molding in capillaries (MIMIC) technique on each glass, poly(methyl methacrylate) (PMMA), polystyrene (PS) and poly(dimethyl siloxane) (PDMS) substrate. The polyelectrolyte multilayer provides adhesion force via strong electrostatic attraction between cell and surface. On the other hand, PEG microstructures also lead to prevent non-specific binding of cells because of physical and biological barrier. The characteristic of each modified surface was examined by using static contact angle measurement. The modified surface onto several substrates provides appropriate environment for cellular adhesion, which is essential technology for cell patterning with high yield and viability in the micropatterning technology. The proposed method is reproducible, convenient and rapid. In addition, the fabrication process is environmentally friendly process due to the no use of harsh solvent. It can be applied to the fabrication of biological sensor, biomolecules patterning, microelectronics devices, screening system, and study of cell-surface interaction.

• Journal of Dental Rehabilitation and Applied Science
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• v.18 no.2
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• pp.73-80
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• 2002

치과용 임플란트의 세라믹 지대주를 위해 개발된 지르코니아 함유 세라믹 시편의 생체 적합성을 평가하기 위하여 시험관내 세포 독성을 검사를 시행하였다. L929 섬유모세포를 $37^{\circ}C$, 90% 습도, 5% CO2 및 95% 공기의 조건을 유지하는 세포 배양기에서 배양하여 실험에 사용하였다. 배양 2일, 4일, 6일 마다 시편을 넣은 배양 접시 내의 전체 세포 수와 생존 세포 수를 세어 세포 증식과 세포 생존율 검사를 시행하였다. millipore filter test를 이용하여 succinate dehydrogenase 효소 활성을 검사하였으며, 세포막 투과성의 변화를 관찰하기 위해 agar overlay test를 시행하였다. 음성 대조군은 시편을 사용하지 않았으며, 양성 대조군은 시편과 같은 크기의 구리를 사용하여, 다음과 같은 결과를 얻었다. 1. 세포 증식과 세포 생존율 검사에서는 지르코니아 함유 세라믹을 넣은 실험군과 음성 대조군 모두에서 시간이 경과함에 따라 세포가 증식하는 양상을 보였다. 세포 생존율 검사에서도 실험군과 음성 대조군이 유사한 결과를 나타내었다. 2. millipore filter test에서는 실험 시편 모두에서 염색 정도의 변화가 없이 음성 대조군과 동일한 결과를 나타냈다. 반면에 구리 시편을 넣은 양성 대조군에서는 중등도의 세포 독성을 나타냈다. 3. agar overlay test에서도 시편을 넣지 않은 음성 대조군에서는 세포 성장에 변화가 나타나지 않았으며, 실험군에서도 시편 주위로 탈색이 관찰되지 않아서 음성대조군과 같은 결과를 나타냈다. 양성 대조군에서는 심한 세포 독성을 나타내었다. 4. 실험결과, 치과용 임플란트의 세라믹 지대주를 위해 개발된 지르코니아 함유 세라믹 시편은 시험관내 세포 독성을 나타내지 않았다.

• Korean Journal of Animal Reproduction
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• v.20 no.3
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• pp.279-288
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• 1996

The objective of this study was to determine the effect of EGF on the development of IVM/IVF bovine embryos and their ICM and TE cell number. In addition, we examined the combined effect of EGF and coculture to the bovine embryo development and the expression of EGF-R protein on bovine embryos by indirect immunofluorescence. The results obtained in these experiments were summarized as follows: When the IVM/IVF 4- to 8-cell embryos were treated at 0, 1, 10, 100 ng/ml of EGF, EGF treatment group showed improved development to blastocyst and increased pattern of ICM and TE cell number compared with control, although there is not significantly different. The stimulating effect of EGF (10 ng/ml) to the develop ment level of IVM/IVF bovine embryos significantly increased development rate to blastocyst after 8-cell stage (p<0.05), although there is no significant effect to the increase of ICM and TE cell numbers. Also, expression of EGF-R on the bovine embryonic stage by indirect immunofluor escence presents after 4-cell stage and the intensity of the EGF-R staining was variable with the development progression. On the other hand, embryos cultured in coculture group added either with or without EGF commonly indicated the significant difference in development rate to blastocyst and Total cell number compared with control. These results suggest that the a addition of EGF to the coculture may stimulate the coculture effect between IVM/IVF bovineembryos and cumulus cells. Therefore, EGF could promote preimplantation bovine embryo development by binding with expressed EGF~R after 4-cell stage, and stimulate the production of embryotrophic factors from the coculture environment. Also, the present study showed that there was no significant effect of EGF to the increase of ICM and TE cell number although the rate of blastocyst significantly increased when treated with EGF after 8-cell stage (p<0.05).

• Restorative Dentistry and Endodontics
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• v.34 no.5
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• pp.430-441
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• 2009

The purpose of this study was to characterize functional distinction between human dental pulp cells(PC) and periodontal ligament cells(PDLC) using cDNA micro array assay and to confirm the results of the microarray assay using RT-PCR. 3 genes out of 51 genes which were found to be more expressed(>2 fold) in PC were selected, and 3 genes out of 19 genes which were found to be more expressed(>2 fold) in PDLC were selected for RT-PCR as well. According to this study, the results were as follows: 1. From the micro array assay, 51 genes were more expressed (2 fold) from PC than PDLC. 2. RT-PCR confirmed that ITGA4 and TGF ${\beta}2$ were more expressed in PC than in PDLC 3. From the micro array assay, 19 genes were more expressed (2 fold) from PDLC than PC. 4. RT-PCR confirmed that LUM, WISP1. and MMP1 were more expressed in PDLC than in PC. From the present study, different expression of the genes between the PC and PDLC were characterized to show the genes which play an important role in dentinogenesis were more expressed from PC than PDLC, while the genes which were related with collagen synthesis were more expressed from PDLC than PC.