• Development and Reproduction
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• v.15 no.1
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• pp.41-52
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• 2011

In the present study, embryoid bodies (EBs) obtained from induced pluripotent stem cells (iPSCs) were induced to differentiate into germ lineage cells by treatment with bone morphogenetic protein 4 (BMP4) and retinoic acid (RA). The results were compared to the results for embryonic stem cells (ESCs) and multipotent spermatogonial stem cells (mSSCs) and quantified using immunocytochemical analysis of germ cell-specific markers (integrin-${\alpha}6$, GFR-${\alpha}1$, CD90/Thy1), fluorescence activating cell sorting (FACS), and real time-RT-PCR. We show that the highest levels of germ cell marker-expressing cells were obtained from groups treated with 10 ng/$m{\ell}$ BMP4 or 0.01 ${\mu}M$ RA. In the BMP4-treated group, GFR-${\alpha}1$ and CD90/Thy-1 were highly expressed in the EBs of iPSCs and ESCs compared to EBs of mSSCs. The expression of Nanog was much lower in iPSCs compared to ESCs and mSSCs. In the RA treated group, the level of GFR-${\alpha}1$ and CD90/Thy-1 expression in the EBs of mSSCs Induced pluripotent stem cells, Mouse embryonic stem cells, Multipotent spermatogonial stem cells, Germ cell lineage, Differentiation potential. was much higher than the levels found in the EBs of iPSCs and similar to the levels found in the EBs of ESCs. FACS analysis using integrin-${\alpha}6$, GFR-${\alpha}1$, CD90/Thy1 and immunocytochemistry using GFR-${\alpha}1$ antibody showed similar gene expression results. Therefore our results show that iPSC has the potential to differentiate into germ cells and suggest that a protocol optimizing germ cell induction from iPSC should be developed because of their potential usefulness in clinical applications requiring patient-specific cells.

• Development and Reproduction
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• v.5 no.1
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• pp.53-57
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• 2001

Sperm capacitation and acrosome reaction (AR) have been known to be Ca$^{2+}$-dependent events. Sperm capacitation accompanies with cholesterol efflux fiom plasma membrane, that eventually stimulates AR. However, whether the AR mediated by cholesterol efflux is Ca$^{2+}$ dependent has not been verified yet. Recently, methyl beta cyclodextrin (MBCD) was found to evoke AR by stimulating the cholesterol efflux fiom sperm membrane. In the present study, we examined the requirement of Ca$^{2+}$ in the MBCD-induced AR. During incubation of sperm in the bicarbonate buffered media MBCD increased AR in a dose-dependent manner regardless of the Ca$^{2+}$ presence. In the presence of low molar concentration of Ca$^{2+}$ (100 ${\mu}$M), MBCD-induced AR was slightly increased compared to Ca$^{2+}$-free condition. In the absence of Ca$^{2+}$ supplement, spontaneous AR was slightly increased during the incubation but inhibited by 100 ${\mu}$M EGTA. MBCD potentiated AR even the presence of EGTA. However, EGTA attenuated MBCD-induced AR, suagesting the functional involvement of intracellular Ca$^{2+}$ in the MBCD-induced AR. Taken together, it was suggested that cholesterol efflux from the sperm plasma membrane was sufficient for induction of AR even in the absence of extracellular Ca$^{2+}$and that a condition permissive for mobilization of intracellular Ca$^{2+}$ is important for MBCD-induced AR.

• Food Science and Preservation
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• v.19 no.5
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• pp.792-798
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• 2012

The purpose of this study was to evaluate the anti-atopic-dermatitis effect of Angelica keiskei extract using a DNFB-induced animal model of atopic-dermatitis symptoms. A. keiskei was prepared via extraction with DW, 50% ethanol, and 100% ethanol in addition to fresh juice. All the extracts reduced the thickness levels of the ears and ear epidermis against swelling by DNFB inducement, especially in the aqueous-extract-treated group. The mRNA expressions of matrix metalloproteinases (MMPs), however, were not observed, which may indicate that Angelica keiskei extract alleviates the atopic-dermatitis symptoms in an MMP-independent manner. Finally, the level of the inflammatory cytokine IL-4 was inhibited only in the juice-treated group, although the IL-13 level was inhibited in the juice-, 50%-ethanol-extract-, and 100%-ethanol-extract-treated groups in a dose-dependent manner, and was not inhibited in the aqueous-extract-treated group. Taken together, these results suggest that Angelica keiskei extract has an inhibitory effect on atopic dermatitis, and may be a useful biomaterial for the development of cosmeceuticals.

• Journal of Periodontal and Implant Science
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• v.28 no.2
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• pp.235-247
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• 1998

Calcium sulfate has a long history of medical use as an implant material. The biocompatibiliry of the material has been clearly established. Bone ingrowth concomitant with resorption occurs rapidly with efficient conduction of bone from particle to particle. Calcium sulfate also has a potential for functioning as a good bamer membrane. The purpose of this study was to compare the biocompatibility of different types of calcium sulfate grafting materials including an expelimental calcium sulfate compound on periodontal ligament cells in vitro as a preliminary test towards the development of a more convenient and useful form of grafting material which could promote regeneration of periodontal tissue. Human periodontal ligament cells were collected from the premolar teeth extracted for orthodontic treatment. cells were cultured in a.MEM culture medium containing 20% FBS, at $37^{\circ}C$ and 100% humidity, in a 5% CO2 incubator. Cells were cultured into 96 well culture plate $1{\times}104$ cells per well with $\alpha$-MEM and incubated for 24 hours. After discarding the medium, those cells were cultured in $\alpha$-MEM contained with 10% FBS alone (control group), in medcal-grade calcium sulfate(MGCS group), in plaster(plaster group), experimental calcium sulfate paste(CS paste group) for 1, 2, 3 day respectively. And then each group was characterized by examining of the cell counting, MTI assay, collagen synthesis. The results \vere as follows. 1. In the analysis of cell proliferation by cell counting, both medical-grdde calcium sulfate group and plaster group showed no stastically significant difference at day 1, 2, 3 accept for plaster group at day 1 compared to control group, but there was stastically significant difference between CS paste group and all other groups at day 1, 2, 3(P<0.05). 2. In the analysis of cytotoxicity by MIT assay, both medical-grade calcium sJlfate group and plaster group showed no stastically significant difference compared to control group at day 1, 2, 3 but there was stastically significant difference between CS paste group and all other groups at day 1, 2, 3(P<0.OS). 3. In the analysis of collagen synthesis by immunoblotting assay, high level was detected for medical-grade calcium sulfate group and plaster group at day 1, 2, 3 compared to CS paste group. On the basis of these results, medical-grade calcium sulfate and plaster was shown to possess biocompatibility whereas the CS paste had unfavourable outcome. This observation shows a need for modification of the materials contained in calcium sulfate paste.

• Journal of Periodontal and Implant Science
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• v.31 no.2
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• pp.401-420
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• 2001

A novel glucanhydrolase from a mutant of Lipomyces starkeyi(KSM 22)has been shown effective in hydrolysis of mutan, reduction of mutan formation by Streptococcus mutans and removal pre-formed sucrose-dependent adherent microbial film and Lipomyces starkeyi KSM 22 dextranase has been strongly bound to hydroxyapatitie. These in vitro properties of Lipomyces starkeyi KSM 22 dextranase are desirable for its application as a dental plaque control agent. This study was performed to determine oral hygiene benefits and safety of dextranase(Lipomyces starkeyi KSM 22 dextranase)-containing mouthwash in human experimental gingivitis. This 3-week clinical trial was placebo-controlled double-blind design evaluating 1U/ml dextranase mouthwash and 0.12% chlorhexidine mouthwash. A total 39 systemically healthy subjects, who had moderate levels of plaque and gingivitis were included. At baseline, 1, 2 and 3 weeks, subjects were scored for plaque(Silness and $L{\ddot{o}e$ plaque index and plaque severity index), gingivitis($L{\ddot{o}e$ and Silness gingival index), and at baseline and 3 weeks of experiment, subjects were scored for plaque(Turesky-Quingley-Hein's plaque index and plaque severity index), tooth stain(Area and severity index system by Lang et al). Additionally, oral mucosal examinations were performed and subjects questioned for adverse symptoms. Two weeks after pre-experiment examinations and a professional prophylaxis, the subjects provided with allocated mousewash and instructed to use 20-ml volumes for 30s twice dailywithout toothbrushing. All the groups showed significant increase in plaque accumulation since 1 week of experiment. During 3 weeks' period, the dextranase group showed the least increase in plaque accumulation of Silness and $L{\ddot{o}e$ plaque index, compared to the chlorhexidine and placebo groups, but chlorhexidine group showed the least increase inplaque accumulation of Turesky-Quingley-Hein's plaque index. As for gingival inflammation, all the groups showed significant increase during 3 weeks of experiment. The dextranase group also showed the least increase in gingival index score, compared to the chlorhexidine as well as the placebo groups. Whereas the tooth stain was increased significantly in the chlorhexidine group, compared to the baseline score and the placebo group since 3 weeks of mouthrinsing. It was significantly increased after 3 weeks in the dextranase group, still less severe than the chlorhexidine group. As for the oral side effect, the dextranase group showed less tongue accumulation, bad taste, compared to the chlorhexidine group. From these results, mouthrinsing with Lipomyces starkeyi KSM 22 dextranase was comparable to 0.12% chlorhexidine mouthwashin inhibition of plaque accumulation and gingival inflammation and local side effects were if anything less frequent and less intense than chlorhexidine, in human experimental gingivitis. All data had provided positive evidence for Lipomyces starkeyi KSM 22 dextranase as an antiplaque agent and suggested that further development of dextranase formulations for plaque control are warranted.

• Journal of Life Science
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• v.17 no.11
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• pp.1492-1496
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• 2007

In vitro IgE class switching could be induced through co-culture of CD40L-expressing KU812 cells and CD40-expressing B cells in the presence of IL-4 or IL-13. It has been generated several B cell lines, which produce rice allergen (RA)-specific IgM antibody by in witγo immunization (IVI) using peripheral blood lymphocyte (PBL). In this study, induction of RA-specific IgE antibody by KU812 cells was attempted. Before co-culture, we determined the CD40 expression in RA-specific B cell lines, RA9G11 and the CD40 ligand (CD40L) expression in activated KU812 cells by treatments with phorbol myristate acetate (PMA) and ionomycin for 6 hrs. Flow cytometric analysis shown that RA9G11 and activated KU812 cells expressed high level of CD40 and CD40L, respectively. RA9G11 cells were cultured with activated KU812 cells for 12 days in the presence of IL-4 for IgE class switching. Mature $C{\varepsilon}$ mRNA level and RA-specific IgE spot forming cells (SFC) were observed in all culture condition, and especially, high level of RA-specific IgE synthesis was determined the same ratio of RA9G11 and activated KU812 cells in the presence of 50U IL-4. Therefore, induction of RA-specific IgE synthesis by activated KU812 cells can be contributed in the application for allergic therapy and prevention.

• Journal of Life Science
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• v.33 no.6
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• pp.512-522
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• 2023

Cancer with unlimited cell growth is a leading cause of death globally. Various cancer treatments, including surgery, chemotherapy, radiation therapy, immunotherapy, and targeted therapy, can be applied alone or in combination depending on the cancer type and stage. New treatments with fewer side effects than previous cancer treatments are continually under development and in demand. Undifferentiated stem cells with unlimited cell growth are gradually changed via cellular differentiation to arrest cell growth. In this study, we reviewed the possibility of treating cancer by using cellular differentiation into the adipocytes in cancer cells. In previous in vitro studies, oral antidiabetic drugs of the thiazolidinedione (TDZ) class, such as rosiglitazone and pioglitazone, were induced into the adipocytes in various cancer cell lines via increased peroxisome proliferator-activated receptor-γ (PPAR γ) expression and glucose uptake, which is the key regulator of adipogenesis and the energy metabolism pathway. The differentiated adipogenic cancer cells treated with TDZ inhibited cell growth and had a less cellulotoxic effect. This adipogenic differentiation treatment suggests a possible chemotherapy option in cancer cells with high and abnormal glucose metabolism levels. However, the effects of the in vivo adipogenic differentiation treatment need to be thoroughly investigated in different types of stem and normal cells with other side effects.

• Journal of Embryo Transfer
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• v.24 no.3
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• pp.199-205
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• 2009

This study was carried out to investigate the effective genetic resources preservation system using the frozen boar semen. The porcine oocytes were matured for 44 hours in NCSU-23 medium with or without 10% Porcine Follicle Fluid (PFF), 0.5 ${\mu}g/ml$ porcine FSH, 0.5 ${\mu}g/ml$ equine LH, 1.0 ${\mu}g/ml$ 17 $\beta$-estradiol ($E_2$) and 10 ng/ml Epidermal Growth Factor (EGF) under mineral oil at $38.5^{\circ}C$ in humidified atmosphere of 5% $CO_2$ in air. After 44 h of culture, the oocytes were inseminated with frozen-thawed semen and fresh semen prepared with mTBM medium for 6 h. Later, set of 50 presumptive zygotes were transferred into 4-well dish (500 ${\mu}l$) of IVC medium. for embryos freezing, slow-freezing and vitrification methods were used as a cryopreservation. Differences among treatments were analyzed using General Linear Model Procedure by SAS Package (version 6.12) differences were considered significant when p<0.05. Following IVF and IVC, the rates of cleavage and blastocysts formation were significantly higher (p<0.05) in hormone supplemented group than that of hormone-free group (25.7 vs, 12.1). The development rates to cleavage and blastocysts were significantly higher in PZM-5 group than NCSU-23 group (60.3%, 46.6% vs 27.4%, 11.1%). Further improvement was achieved when PZM-5 was supplemented with FBS. Cleavage rates was significantly higher in fresh semen source group than frozen semen (66.7% vs 43.7%). However in blastocysts rates was similar two groups. Post-thaw survival rates of embryos were 1.2% and 2.2% in slow-frezing and vitrification groups, respectively. The results of our study suggest that it is still possible to improve the culture conditions and boar semen cryopreservation for enhance reproductive technology and animal genetic resources conservation.

• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.6
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• pp.1061-1068
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• 2011

Moxibustion is a treatment method for cold and pain in Korean medicine. But the systematic study of moxibustion were very few. This study was aimed to survey and evaluate moxibustion study in Korean medicine. We search the moxibustion study in journals related Korean medicine and http://oasis.kiom.re.kr. The period of the study from first issue to August, 2010. The search word were moxibustion, moxa in keyword and moxibustion treatment in title. We totally search 334 articles. But 175 articles were excluded because this study were not exactly moxibustion study, so we included and analyzed 159 articles. The study of moxibustion were 159 articles. Literature review were 35 articles, systematic review 8, heat experiment 10, in vivo or vitro 28 and survey investigation 3. Clinical articles of moxibustion related to Korean medical journals were 76. Before and after study were 31 articles, case report 20, cohort study 7, cross-sectional study 2, nonrandomized study 6, quasi randomized study 1, randomized clinical trials 9. This results showed that moxibustion study were smaller than acupuncture or herb and concentrated to pain and cold syndrome. We need further larger and diverse study of moxibustion.

• Journal of Life Science
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• v.28 no.7
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• pp.827-834
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• 2018

In this study, the biological activities of aqueous, ethanol, and methanol extracts of larvae of the edible insect Protaetia brevitarsis seulensis, fermented using several kinds of microorganisms, were tested in in vitro experimental models. Six effective microorganisms were used for fermentation, namely Lactobacillus plantarum JBMI F3, Lactobacillus plantarum JBMI F5, Lactobacillus gasseri Ba9, Aspergillus kawachii KCCM 32819, Saccharomyces cerevisiae KACC 93023, and Bacillus subtilis KACC 91157. Biological activities (${\alpha},{\alpha}^{\prime}-diphenyl-{\beta}-picrylhydrazyl$ [DPPH] free radical scavenging activity, reducing power, and fibrinolytic activity), and biochemical properties (phenolic compounds and flavonoids) were examined in aqueous, ethanol, and methanol extracts from P. brevitarsis seulensis powder and fermented P. brevitarsis seulensis powder. The total phenolic compounds and flavonoid contents were highest in the aqueous extract of B. subtilis-fermented P. brevitarsis seulensis powder. DPPH radical scavenging activity and reducing power were stronger in the fermented group than the nonfermented group. Fibrinolytic activity were highest in the extract from B. subtilis-fermented P. brevitarsis seulensis powder. The ${\alpha}-amylase$ activity in starch was higher in the fermented group than the nonfermented group, but there was no significant difference. These results provide basic data to understand the biological activities of bioactive materials derived from fermented P. brevitarsis seulensis larvae for the development of functional foods.