• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.6
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• pp.564-570
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• 2005

In the present study, the protective effects of Bcl-2 over-expression in a suspension culture (without any adaptation) and spent medium (low nutrient and high toxic metabolite conditions) were investigated. In the suspension culture without prior adaptation, the viability of the control cell line fall to 0% by day 7, whereas the Bcl-2 cell line had a viability of 65%. The difference in the viability and viable cell density between the Bcl-2 and control cell lines was more apparent in the suspension culture than the static culture, and became even more apparent on day 6. Fluorescence microscopic counting revealed that the major mechanism of cell death in the control cell line in both the static and suspension cultures was apoptosis. For the Bcl-2 cell lines, necrosis was the major mode of cell death in the static culture, but apoptosis became equally important in the suspension culture. When the NS0 6A1 cell line was cultured in spent medium taken from a 14 day batch culture, the control cell line almost completely lost its viability by day 5, whereas, the Bcl-2 still had a viability of 73%. The viable cell density and viability of the Bcl-2 cell line cultivated in fresh medium were 2.2 and 2.7 fold higher, respectively, than those of the control cultures. However, the viable cell density and viability of the Bcl-2 cultivated in the spent medium were 8.7 and 7.8 fold higher, respectively, than those of the control cultures. Most of the dead cells in the control cell line were apoptotic; whereas, the major cell death mechanisms in the Bcl-2 cell line were necrotic.

• Journal of Korean Academy of Oral and Maxillofacial Radiology
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• v.25 no.2
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• pp.331-341
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• 1995

The purpose of this study was to aid in the prediction of tumor cell tolerance to radiotherapy and/or chemotherapy. For this study, cell surviving curves were obtained for murine melanoma Bl6 cell line using semiautomated M1T assay. 2,4,6,8, 10Gy were irradiated at a dose rate of 210cGy/min using /sup 60/Co Irradiator ALOORADO 8. After irradiatior, B16 cell lines(2.5×10⁴ cells/ml) were exposed to bleomycin and cisplatin at concentration of 0.2㎍/㎖, 2㎍/㎖ and 20㎍/㎖ for I hour respectively. The viable cells were determined for each radiation dose and/or each concentration of drug. And they were compared to control values. The obtained results were as follows : 1. There was significant difference of surviving fraction at 4, 6, 8, 10Gy on B16 cell line(P<0.05). 2. There was significant difference of cytotoxicity between bleomycin and cisplatin at concentration of 0.2㎍/㎖ and 2㎍/㎖(P<0.05) on B16 cell line, but there was no significant difference of cytotoxicity at concentration of 20㎍/㎖ on B16 cell line. 3. There was significant difference of cytotoxicity of bleomycin after irradiation of 2Gy and 10Gy on B16 cell line(P<0.01). 4. There was significant difference of cytotoxicity of cisplatin at concentration of 20㎍/㎖ after irradiation on B16 cell line.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.4 no.1
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• pp.177-197
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• 1998

This experimental study was carried out to evaluate the effect of Yipahnsan on angiogenic inhibition mechanism. This study investigates the effects of Yipahnsan on angiogenic inhibition mechanism evaluate cell adhesive inhibition effect, DNA fragmantaion analysis, nuclear condensation assay, FACScan analysis, angiogenic lumen formation assay, immunocytochemistry analysis, RT-PCR for mRNA expression, western blot analysis, confocal analysis for $Ca^{2+}influx$. The results were summarized as follows : 1. The cell adhesive inhibition ability was strongly increased from $5{\mu}g/ml$ on ECV304 cell line and ECVPAR cell line. 2. YY water extract caused $G_0/G_1$ arrest peak to existed on the ECV304 cell line. 3. YY water extract caused inhibition of proliferation and inducement of apoptosis on the collagen coated plate in ECV304 cell line. 4. YY water extract inhibited the lumen formation on the matrigel coated plate in ECV304 cell line. 5. YY water extract inhibited the expressions of LFA-1 and ELAM-1 on ECV304 cell line and ECVPAR cell line. 6. YY water extract inhibited the expressions of MMP-9 and uPA on ECV304 cell line and ECVPAR cell line. 7. YY water extract inhibited the expression of integrin ${\alpha}_v{\beta}_3$ on ECV304 cell line and ECVPAR cell line. 8. YY water extract decreased the change of $Ca^{2+}$ in intracellular on ECV304 cell line and ECVPAR cell line. According to the results, Yipahnsan showed to be a key antagonist of integrin ${\alpha}_v{\beta}_3$, and to be induction of apoptosis by p53 through flow cytometry. This report also demonstrated that expressions of MMP-9 and uPA were blocked under the angiogenesis model. Thus, we suggested that Yipahnsan blocks angiogenesis by inducing apoptosis in ECV304 and ECVPAR cell lines, and another oriental herbal medicine that treats qi-stagnation and blood-stasis type also has angiogenic inhibition effects.

• Journal of Nutrition and Health
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• v.31 no.4
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• pp.799-808
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• 1998

To investigate the antitumor activity of mugwort (Artemisia princeps Pampan), petroleum ether extract of mugwork was partially purified by a silica gel chromatography. Among several fractions, the fraction which was obtained under the elution with acetone, showed potent cytotoxicity against mouse leukemia cell line(Ll210), human colon cancer cell line (HCT-48) and human hepatoma cell line (Hep G2) , but was less effective with normal cell line(mouse embryo cell). Acetone fraction appeared to be glycolipid by Benedict test and the major fatty acids of the lipid were C16 ; 0 , C 18: 3by GC/MS analysis.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.10
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• pp.1338-1343
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• 2004

The Debao pony ear marginal tissue fibroblast cell line (NDPEM 2/2) was uccessfully established using either primary explant technique or collagenase technique. The characterizations of the cell line were identified as following: the cells were adherent and of density limitation; population doubling time (PDT) of cells made with the two techniques were 35.9 h and 48 h, respectively; chromosome analysis showed that the frequency of cell chromosome number to be 2n=64 was 91.3%-92.8%. Confirmed by isoenzyme analysis, this cell line had no cross- contamination. Tests for microbial contamination from bacteria, fungi, virus or mycoplasma were negative. This newly established cell line meets all the standard quality controls of ATCC. It will provide a precious genetic resource for the conservation of the Debao pony breed, as well as effective experimental material for genetic studies on Debao ponies.

• Advances in Traditional Medicine
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• v.3 no.3
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• pp.151-156
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• 2003

The leaves of Mirabilis jalapa L contains protein fraction presumed ribosome-inactivating protein (RIP). RIP is a group of protein that has RNA N-glycosidase activity that is capable to inhibit protein synthesis. Protein fraction of the plant was shown to be cytotoxic on HeLa cell-line, however, the mechanism by which the protein kill the cells is not identified yet, whether trough apoptosis, necrosis, or other mechanism. This research aim to study the mechanism of cell death caused by the protein fraction isolated from the leaves of this plant on HeLa and Raji cell-line, as representative of different kind of cancer cells. Results showed that protein fraction isolated from the leaves of Mirabilis jalapa L was more cytotoxic to HeLa cell-line (LC50: 0.65 mg/ml) than to Raji cell-line (1.815 mg/ml) on 48 hours incubation time. Moreover, it was demonstrated that the death of HeLa cells caused by the protein fraction was due to induction of apoptosis, while on Raji cell-line was due to non-apoptosis way, presumably via necrosis.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.66-69
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• 2001

Previously we developed a CHO cell line (CHO-OTC1-A19) expressing the first two enzymes of urea cycle. This cell line showed higher ammonia removal activity and faster growth rate than the vector controlled CHO cells (CHO-neo-5). The purpose of this study was to develop a cell line with higher ammonia removal activity than the cell line developed previously. To accomplish this, we constructed stable CHO cell lines expressing the first three, the first four, or all five enzymes of urea cycle by the stable transfection method. We finally selected CHO-AL-19 cell line expressing the first three, the first four enzymes of the cycle with higher ammonia activity than CHO-OTC1-A19 and CHO-n대-5 cell lines: 40% and 15% higher than those of CHO-neo-5 and CHO-OTC1-A19 cell lines 72 hour after culture started, respectively. It also showed 44% and 10% higher cell viability than CHO-neo-5 and CHO- OTC1-A19 cell lines at higher cell density. In addition, CHO-AL-19 cells showed 45%-60% and about 20% lower ammonia concentration per cell than those of CHO-neo-% and CHO-OTC1-A19 cell lines, respectively. These results indicate that CHO-AL-19 could be used in the production of human therapeutic proteins with higher efficiency.

• Journal of Korean Academy of Oral and Maxillofacial Radiology
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• v.27 no.1
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• pp.43-53
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• 1997

The purpose of this study was to aid in the prediction of tumor cell tolerance to radiotherapy and/or chemotherapy. For this study, cell surviving curves were obtained for mouse lymphoma YAC-1 cell line using semi automated MTT assay. 2, 4,6, 8, 10Gy were irradiated at a dose rate of 210cGy/rnin using /sup 60/Co Irradiator ALDORADO 8. After irradiation, YAC-1 cell lines(3×10⁴cells/ml) were exposed to bleomycin or cisplatin for 1 hour. The viable cells were determined for each radiation dose and/or each concentration of drug at the 4th day. And they were compared to control values. The obtained results were as follows : 1. The surviving curve with gentle slope was obtained after irradiation of 2, 4, 6, 8, 10 Gy on YAC-1 cell line. 2. The cytotoxicity of bleomycin or cisplatin was increased significantly at all concentration of 0.2㎍/ml, 2㎍/ml and 20㎍/ml on YAC-1 cell line (P<0.01). And the cytotoxicity of cisplatin was greater than that of bleomycin at all concentration on YAC-1 cell line (P<0.01). 3. There were no significant differences of surviving fractions among 4Gy, 6Gy and 8Gy after irradiation of each radiation dose with 2㎍/ml of bleomycin compared with irradiation only on YAC-1 cell line. 4. There was significant difference of surviving fraction between 2Gy and 10Gy after irradiation of each radiation dose with 2㎍/ml of cisplatin compared with irradiation only on YAC-1 cell line(P<0.05). 5. There were significant differences of surviving fractions between the groups of irradiation only and the groups of irradiation with 2㎍/ml of bleomycin or cisplatin at all doses of 2, 4, 6, 8 and 10Gy on YAC-1 cell line(P<0.05).

• Journal of Korean Academy of Oral and Maxillofacial Radiology
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• v.24 no.2
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• pp.249-260
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• 1994

The purpose of this study was to aid in the radiation therapy of head and neck cancer patients. For this study, radiation survival curves were generated for Bl6, MG-63 and YAC-l cell lines using semiautomated MTT assay and Dye Exclusion Assay. Irradiation of 2, 4, 6, 8, 10Gy were delivered at room temperature at a dose rate of 210.2cGy/min using / sup 60/Co γ-ray Irradiator ALOORAOO 8. The viable cells were determined for each radiation dose and compared to control values. The obtained results were as follows: 1. There was significantly different absorbance at 10Gy on B16 cell line in MTT assay(P<0.05). 2. There was significantly different absorbance at 4, 6, 8, 10Gy on MG-63 cell line in MTT assay(P<0.05). 3. YAC-l cell line was more sensitive than B16 or MG-63 cell line to all doses of radiation(P<0.05). 4. There was significantly different absorbance among all tumor cell lines except between B16 and MG-63 cell line at ZGy in MTT assay(P<0.05). 5. Good correlation was obtained between MTT assay and DEA(P<0.05). The efficient of correlation of B16, MG-63 and YAC-l cell line was 0.845, 0.824 and 0.906, 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.