• Journal of Life Science
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• v.5 no.1
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• pp.1-7
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• 1995

Mammary orgamoids(ductal and endbud fragments) were cultured in a complete hormone medium(CHM) with 10%FBS, estradiol, progesterone, hydrocortisone, insulin, and prolactin, Several types of colonies were observed: stellate(14$$\pm$5.5%), duct(41$\pm$5.6%), web(35$\pm$3.6%), squamous(6$\pm$2.1%), and lobuloduct(4$\pm$1.2%), Squamous colony was typical squamous metaplasia(SM) with several layers of squamous epithlia and keratin pearls. At the immunocytochemical study, casein proteins were predominantly localized near the apical surfaces of the cells or in the lumina of ductal or lobuloductal colonies. To inhibit the formation of SM, we treated organoids with all-trans retinoic acid(RA) from 10$^{-6}$ to 10$^{-17}$ M in CHM. Formation of SN was completely inhibited at 10$^{-9}$M RA in CHM. The frequency of lobuloductal colony formation was increased with the augmentation of RA concentration.

• Restorative Dentistry and Endodontics
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• v.26 no.4
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• pp.285-295
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• 2001

The objective of this study was to investigate the inhibitory effect of taurine and alendronate on the osteoclast differentiation. Osteoblasts and bone marrow cells from 1-2 day old mouse were co-cultured in 10% fetal bovine serum - minimal essential media (FBS-MEM). Osteoclast differentiation was induced by adding the sonicated extracts of Porphyromonas gingivalis (P.gingivalis). Osteoclasts were identified using tartrate resistant acid phosphotase staining (TRAP). Alendronate of 10$^{-7}$, 10$^{-6}$, 10$^{-5}$M and taurine of 500, 1000, 1500$\mu\textrm{g}$/ml were added respectively. The cytotoxic effects of alendronate and taurine were examined using MTT(3-(4,5-dimethylthiazol -2-yl-2,5-diphenyltetrazo- lium bromide) method. After culturing with the sonicated extracts of P.gingivalis, the amounts of IL-6 in the culture supernatant were measured and compared using the ELISA method. The results were as follows : 1. Osteoclasts were differentiated at the concentration of 0.01~0.1$\mu\textrm{g}$/ml sonicated extracts of P.gingivalis. (P<0.05). 2. Alendronate inhibited osteoclasts differentiation at the concentration of 10$^{-5}$ M when the concentration of sonicated extracts of P.gingivalis was 0.01$\mu\textrm{g}$/ml. 3. Taurine inhibited osteoclasts differentiation at the concentration of 1500$\mu\textrm{g}$/ml when the concentration of sonicated extracts of P.gingivalis 0.01$\mu\textrm{g}$/ml. 4. In cytotoxic test (MTT test), no cytotoxic effect was evident in all concentrations of alendronate and taurine. 5. Taurine (10$^{-5}$M) and alendronate(1500$\mu\textrm{g}$/ml) did not change the amounts of IL-6 induced by sonicated extracts of P.gingivalis significantly.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.2
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• pp.358-364
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• 2006

To investigate the possible mechanism of Drynariae Rhizoma extracts as a candidate of anti-cancer drug, I examined the effects of Drynariae Rhizoma extracts on the apoptosis of U937 cell line. MTT assay, flow cytometric analysis, SDS-polyacrylamide gel electrophoresis, Western blot analysis, and RT-PCR were performed. Drynariae Rhizoma extracts treatment reduced the cell viablilty of U937 cells in a dose-dependent manner, which was associated with induction of apoptotic cell death. Drynariae Rhizoma extracts treatment also reduced the levels of Bcl-xL anti-apoptotic protein expression and increased the levels of caspase-3, p53, pro-apoptotic protein, in U937 cells. RT-PCR data revealed that the level of bcl-2, bcl-xL mRNA expressions decreased in a dose-dependent manner. These findings suggest that Drynariae Rhizoma extracts may have induction of apoptotic cell death via regulation of several growth regulatory gene products. The abbreviations used are: FBS, fetal bovine serum; PBS, phosphate buffered saline; PI, propidium iodide; OD, optical density; DiOC6, 3,3-dihexyloxa carbcyanine iodide; MTT, 3 [4-5-dimethylthiazol-2-yl] -2-diphenyltetrazolium bromide.

• Biomolecules & Therapeutics
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• v.4 no.1
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• pp.46-50
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• 1996

Inhibition of the growth of human cancer cells by proteins secreted from 373-L1 cells was investigated in the present study. The growth of human cancer cells was inhibited by co-culture with 373-L1 cells under 10% FBS and DME, DME, GIT and serumless medium, respectively. The conditioned medium of cultured 373-L1 cells under serumless medium was concentrated 100-fold through an ultrafiltration cell with a 10,000 molecular weight cutoff at 4$^{\circ}C$ under positive pressure using nitrogen(373-L1 EM). 373-L1 EM inhibited the growth of HeLa, Hep G 2, KHOS-Np, A43l and MCF-7 cells. 3T3-L1 EM was purified with FPLC, DEAE-ion exchange chromatography and phenyl-sepharose chromatography. The major protein of 373-L1 EM has a molecular weight of 66,000-68,000 in SDS-PAGE analysis. The results suggest that the inhibitory activity of 373-L1 EM appears to be due to some protein(m.w.66,000-68,000) secreted by 373-L1 cells.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.20 no.3
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• pp.43-50
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• 2007

Background and Objectives : The aim of this study was to investigate the anti-oxidant and anti-inflammatory effects of the Taglisodog-eum(TSE) extract on the RAW264.7 cell Methods : The RAW264.7 cell was cultured using Dulbecco's modified Eagle's medium(DMEM, USA), including the 10% fetal-bovine serum(FBS; Sigma, USA) in a $37^{\circ}C$, 5% CO2 incubator. Results : The anti-oxidant ability of TSE were dose-dependantly increased. The LPS-induced IKK, iNOS and COX-2 mRNA expression were dose-dependantly decreased in the RAW264.7 cells treated with TSE. $NF-kB$ activation was suppressed. Conclusion : The findings in this study show that TSE has anti-oxidant and anti-inflammatory effects, such as the inhibition of $NF-kB$ activity.

• Korean Journal of Animal Reproduction
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• v.27 no.2
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• pp.103-113
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• 2003

This study was conducted to investigate the optimal condition for produce of large quantity recipient oocytes on porcine cloned embryos. In order to determined the optimum concentration and exposure time of ethanol, $Ca^{2+}$-ionophore, 6-DMAP and cycloheximide, in vitro matured oocytes were activated in TCM-199 containing various chemicals and 15% FBS. The activated oocytes were cultured in Whitten's medium containing 10% FBS at 5% $CO_2$. 1. When the porcine oocytes were activated with the ethanol, the best pronucleus formation, cleavage, and in vitro development rate were obtained in the 10% for 10 minutes, which was significantly higher than all of the other treatment(53.4%, 51.6% and 39.9%, respectively). 2. When the porcine oocytes were activated with the $Ca^{2+}$-ionophore, the pronucleus furmation, cleavage, and in vitro development rate were found significantly higher in the 25$\mu$M fur 2min. treatment than those of other concentration and exposure time(59.7%, 62.2% and 43.9%, respectively). 3. When the porcine oocytes were activated with the 6-DMAP, the best pronucleus formation, cleavage, and in vitro development rate were obtained in the 2mM for 2hr~4.5hr(57.3%, 58.4% and 29.0%, respectively). 4. When the porcine oocytes were activated with the cycloheximide, result showed that pronucleus formation, cleavage, and in vifro development rate were 52.1%, 47.7% and 31.8%, respectively, in the 5$\mu\textrm{g}$/ml for 4hr~6hr treatmrent, which was significantly higher than all of the other treatment. These results suggested that the active condition of porcine oocytes was established by optimum concentration and exposure time among different chemicals for produce of large quantity recipient oocytes.s.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.3
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• pp.287-292
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• 1998

The objective of this study was to confirm whether the vitrification method using EFS35 has detrimental effect for cytoskeleton and chromosome constitution of the mouse oocytes by indirect immunocytochemistry and chromosome analysis. Mouse oocytes were vitrified using EFS35 which consisted of 35% ethylene glycol, 18% ficoll, 0.3 M sucrose and 10% FBS in M2 medium. The results obtained in this experiment were summarized as follows: When the survival rates after being exposed or vitrified in EFS35 were examined, there were not different between two groups (97.7 and 89.3%). Also, when the microtubule morphology and microfilament distribution in vitrified oocytes were examined, normal percentage of two cytoskeleton in vitrified group (95.5 and 100%) was not different from that in control (97.5 and 100%) and exposed group (92.3 and 100%). In addition, the rate of oocytes containing a normal chromosome number in vitrified group (73.5%) after IVF was not different from that in control (79.5%) and exposed group (78.7%). These results indicated that the cytoskeletal morphology and chromosome constitution of mouse oocytes were not affected by cryoprotectant (EFS35) or freezing apd that vitrification methods using EFS35 was suitable for cryopreservation of mouse oocytes.

• Journal of Periodontal and Implant Science
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• v.35 no.3
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• pp.623-634
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• 2005

Human periodontal ligament fibroblast(hPDLF) is very important to cure periodontal tissue because it can be diverged into various cells. This study examined the expression of MMP-1, TIMP-1, periodontal ligament specific PDLs22, Type I collagen, Fibronectin, TIMP-2, telomerase mRNA in a replicative senescence of hPDLF. The periodontal ligament tissue was obtained from periodontally healthy and non-carious human teeth extracted for orthodontic reasons at the Chosun University Hospital of Dentistry with the donors' informed consent. The hPDLF cells were cultured in a medium containing Dulbecco's modified Eagle medium(DMEM, Gibco BRL, USA) supplemented with 10% fetal bovine serum(FBS, Gibco BRL, USA) at 37C in humidified air with 5% $CO_2$. For the reverse transcription-polymerase chain reaction(RT-PCR) analysis, the total RNA of the 2, 4, 8, 16, 18, and 21 passage cells was extracted using a Trizol Reagent(Invitrogen, USA) in replicative hPDL cells. Two passage cells, i.e. young cells, served as the control, and ${\beta}-actin$ served as the internal control for RT-PCR The results of this study about cell morphology and gene expression according to aging of hPDLF using RT-PCR method are as follows: 1. The size of hPDLF was increased with aging and it was showed that the hPDLF was dying in the final passage. 2. PDLs22 mRNA was expressed in young hPDLF of the two, four, and six passage. 3. TIMP-1 mRNA was expressed in young hPDLF of the two and four passage. 4. There was a tendency that MMP-1 mRNA was weakly expressed over eighteen. 5. Type 1 collagen mRNA was expressed in almost all passages, but it was not expressed in the final passage. 6. Fibronectin mRNA was observed in all passages and it was weakly expressed in the final passage. 7. TIMP-2 and telomerase mRNA were not expressed in this study. Based on above results, it was observed that PDLs22, Type 1 collagen, Fibronectin, MMP-1. and TIMP-1 mRNA in hPDLF were expressed differently with aging. The study using the hPDLF that is collected from healthy patients and periodontitis patients needs in further study.

• Journal of Acupuncture Research
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• v.21 no.3
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• pp.29-41
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• 2004

Objective : This study was produced to examine the effects of moxibustion that had been played important role to traditional oriental medical treatment on disease. Recently, it was reported that moxi-tar which is generated in the process of moxibustion as burning combustibles decreased NO and iNOS generation in C6-glioma and RAW 264.7 cells in our lab. Methods : C6-glioma cells were cultured in RPMI 1640 with FBS 10% in CO2 incubator. To study the protective effects of moxi-tar, we observed cell viability, DPPH activity, SOD activity, catalase activity and cell morphology after injury with $H_2O_2$. Results and Conclusions : Moxi-tar increased cell viability about twice as much as that of being injury by $H_2O_2$(moxi-tar $40{\mu}g/m{\ell}$, $H_2O_2$ $500{\mu}M$). And the results of free radical scavenger activity($80{\mu}g/m{\ell}$ : $78.91{\pm}4.4%$), SOD activity and catalase activity($80{\mu}g/m{\ell}$ : 21.6unit/mg protein) were increased by moxi-tar as dose-dependent manner. So we concluded that the effects of moxibustion which is played important role in Oriental medicine, might be free radical scavenger effects induced by moxi-tar. Conclusion : These results indicate that tBHP induces apoptosis through a lipid peroxidation-dependent mechanism and JS exerts the protective effect against the apoptosis by preventing peroxidation of membrane lipids.

• Reproductive and Developmental Biology
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• v.31 no.2
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• pp.103-108
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• 2007

This study was carried out to investigate the effects of cryoprotectants, warming solution and removal of lipid on open pulled straw vitrification (OPS) method of porcine embryos produced by nuclear transfer (NT) of fetal fibroblasts. All solutions used during vitrification were prepared with holding medium consisting of 25 mM Hepes buffered TCM199 medium containing 20% fetal bovine serum (FBS) at $38.5^{\circ}C$. The blastocysts derived from NT with or without lipid were vitrified in each medium of different concentrations of dimethyl sulfoxide (DMSO) and ethylene glycol (EG). Also, blastocysts after cryopreservation were warmed into different concentrations of sucrose in warming solution. The optimal concentrations of cryoprotectants in vitrification solution were 10% DMSO + 10% EG in vitrification solution 1 (VS1) and 20% DMSO + 20% EG in vitrification solution 2 (VS2). The optimal concentrations of sucrose were 0.3 M sucrose in warming solution 1 (WS1) and 0.15 M sucrose in warming solution 2 (WS2). lipid removal from oocytes before NT enhanced the viability of NT embryos after vitrification. Our results show that use of the OPS method in conjunction with lipid removal provides effective cryopreservation of porcine nuclear transfer embryos.