• Korean Journal of Animal Reproduction
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• v.25 no.2
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• pp.191-198
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• 2001

The purpose of this study was to investigate the effects of embryo sources such as in vivo vs. in vitro produced blastocyst, and culture systems on the membrane permeability and viability of bovine blastocyst following GMP vitrification. To produce in vivo embryos, six cows were superovulated by administration of follicle stimulation hormone (FSH) and prostaglandin $F_{2{\alpha}}$(PG $F_{2{\alpha}}$). in vitro embryos were produced by two different culture systems, oviduct co-culture (OCS) and defined culture system (HECM-6; DCS). Ovaries were picked up at a local slaughterhouse and transported to laboratory in 3$0^{\circ}C$ saline within 2 h. Ovaries were washed with same saline three times and then placed in saline on warm plate adjusted at 3$0^{\circ}C$ during aspiration. The blastocysts produced were assigned for membrane permeability and viability following GMP vitrification. The membrane permeability of blastocysts was checked in 0.5 M sucrose solution on warm plate at 35$^{\circ}C$ for 0, 2, 5 and 7 min, respectively. Then the diameters (width and length) of embryo cytoplasms were measured by a eyepiece meter, and they were converted to their volume by 4/3 $\pi$ $r^3$. The blastocysts were cryopreserved by GMP vitrification method, where they were sequentially placed into vitrification solution before being loaded into GMP vessels and immersed into L$N_2$ within 20 to 25 sec. Post-thaw blastocysts were serially washed in 0.25 and 0.15 M sucrose in HM and TCM-199 for 5 min each, and then cultured in TCM 199 supplemented with 10% FCS for 24 or 48 h. The volume change of in vivo blastocyst at 0, 2, 5 and 7 min (100, 37.1, 34.3 and 31.6%) was significantly more shrunk than those of in vitro blastocysts derived from OCS (100, 59.8, 48.9, 47.9%) and DCS (100, 57.2, 47.3 and 46.9%) (P＜0.05). The viability of post-thaw blastocyst derived from in vivo (93.6%) was also significantly different from those in OCS and DCS (81.9 and 83.6%; P＜0.05). In the present culture system, the morphology of embryos produced in vitro was similar to that of in vivo embryos, but the quality in membrane permeability and post-thaw viability showed a big difference from their sources as in vivo or in vitro derived from OCS and DCS. The results indicated that the quality of in vivo embryos in membrane permeability and post-thaw viability was better than those of in vitro embryos derived from OCS or DCS.

• Restorative Dentistry and Endodontics
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• v.27 no.6
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• pp.600-611
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• 2002

본 연구의 목적은 클로로필린이 치주인대 세포 생활력에 미치는 영향을 평가하는데 있다. 치주인대 세포는 건강한 사람에서 발치한 소구치의 치주인대 조직으로부터 채취하여 배양하였다. 비교 기준을 설정하기 위하여, HBSS 내에서 $37^{\ circ}C$로 보관한 치주인대 세포수의 50%가 생존하는데 소요되는 시간을 MTT 분석법에 의해 측정하였다. 그 결과는 6시간이었다. HBSS에 클로로필린 10, 100, 500 nM이 각각 첨가된 3군의 실험 보존액과 F-medium, ViaSpan, Likorol 등 모두 6군의 실험 보존액을 96-well plate에 접종한 후, 치주인대 세포를 동일한 수로 분주하였다. 이를 6시간동안 보관한 후, 각 실험군 보존액에서의 세포 생활력을 MTT 분석법으로 측정하였다 또한, HBSS와 F-medium 및 클로로필린 500 nM이 첨가된 HBSS군의 세포들에 대해 유식세포 계측을 시행하여 각각의 세포주기를 분석하였다. 실험 결과, 클로로필린 500 nM이 첨가된 HBSS에서 보관한 세포들이 가장 높은 생활력을 나타내었으며, 클로로필린이 첨가되지 않은 HBSS에 비해서 유의하게 양호한 세포 생활력 유지 능력을 보였다. 클로로필린으로 처치한 세포들은 클로로필린의 농도가 커짐에 따라 세포 생활력이 증가하는 양상을 나타내었다. 유식세포 계측 결과, HBSS와 F-medium 및 클로로필린 500 nM이 첨가된 HBSS에서 보관한 세포의 77~80%가 G0-G1 단계의 세포 주기로 측정되어, 대부분의 세포들이 안정된 세포 대사 상태를 보이는 것으로 나타났다. 결론적으로, 클로로필린 처치는 치주인대 세포의 생활력 유지에 도움이 되는 것으로 사료된다.

• Environmental Analysis Health and Toxicology
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• v.20 no.1
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• pp.23-28
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• 2005

PAH 위해성 평가의 생체지표 개발을 위하여, benzo[a]pyrene을 인체 간암 세포주인 HepG2세포에 처리하여 암 억제 단백질인 p53 및 관련 단백질의 발현 양상에 대하여 연구하였다. HepG2 세포의 생존력은 benzo[a]pyrene을 노출시킨 군에서 농도가 증가할수록 감소하였다. p53과 인산화 p53의 발현 양상은 benzo[a]pyrene 농도 의존적으로 증가하는 경향을 보였으며, 반면에 아세틸화 p53은 benzo[a]pyrene의 농도가 증가할수록 감소하는 경향을 나타내었다. 세포 주기 조절에 관련된 p21 단백질은 화학 물질 처리에 의해서 p53과 마찬가지로 증가하였으나, CdK4와 Rb 단백질의 발현에는 변화가 없었다. 상관분석 결과 Benzo[a]pyrene 노출, 세포 생존력, p53, 인산화 p53, p21이 서로 높은 상관성을 보였다. 본 연구의 결과는 p53 단백질의 축적이 benzo[a]pyrene 독성에 있어 매우 중요한 현상이며, 이는 선택적인 지표와 함께 p53 이 benzo[a]pyrene과 같은 PAH 계열의 물질의 위해성 평가를 위한 민감한 생체 지표로써 개발될 수 있음을 시사한다.

• Journal of Life Science
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• v.21 no.9
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• pp.1266-1273
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• 2011

Saponin is a main component of ginseng widely known as an oriental traditional medicinal ingredient. A variety of biological effects of saponin has been reported, but its action related to skin regeneration has remained unclear so far. In this study, the effect of saponin on matrix metalloproteinase as well as its antioxidant effect in cell free system was examined in human dermal fibroblasts. First of all, as a result of investigating the effect of saponin on cell viability using MTT assay, it was shown to increase cell viability below 10 ${\mu}g$/ml, but it also showed cytotoxicity above 25 ${\mu}g$/ml. The antioxidant effect of saponin was exerted by inhibition of $H_2O_2$ in addition to reducing power above 1 ${\mu}g$/ml. In particular, saponin showed a protective effect on DNA oxidation. Furthermore, it was observed that saponin activates MMP-2 and increases MMP-1 activity in gelatin and casein zymography analyses, respectively, indicating that saponin could have potential a therapeutic agent for anti-aging and skin regeneration.

• Korean Journal of Microbiology
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• v.37 no.2
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• pp.145-150
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• 2001

Reactivation of human cytomegalovirus (HCMV) from latency is often fatal to immunocompromised individuals. To understand the effect of HCMV on human monocytes where HCMV establishes latency, two human monocyte cell lines at different stages in differentiation, THP-1 and HL-60 were infected with HCMV. While the viability and morphology of HL-60 cells were not significantly affected by HCMV, the viability of THP-1 cells was dramatically decreased by HCMV infection. THP-1 cells infected with HCMV became aggregated and adhered to the surface of culture dishes, probably due to the increased expression of adherence molecules CD11b on the infected THP-1 cells. THP-1 cells established a latent HCMV infection were induced to differentiate by treatment with TPA and hydrocortisone. Recovery of infectious HCMV from the culture supernatant of differentiated THP-1 cells was dependent on the time of induction of differentiation after HCMV infection. Thus, in vitro model of reactivation of HCMV from latently infected monocytes was established.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.521.2-521
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• 1986

3$0^{\circ}C$에서 대수증식기 세포를 42$^{\circ}C$에서 1시간동안 배양시킨 다음 3$0^{\circ}C$로 다시 이동시켰을 때 DNA합성은 Chloramphenicol의 존재하에서도 재 개시되었다. 그리고 Chloramphenicol을 처리하였을 때 세포 생존력은 크게 증가하지 않았고, 세포의 모양은 정상세포보다 더 길어졌다.

• Journal of the Korean Applied Science and Technology
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• v.38 no.3
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• pp.629-637
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• 2021

To develop a safer cosmetic preservative, we carried out a comparative study on characteristics of OD and OD-gal, where OD-gal was synthesized from OD using E. coli β-gal. OD-gal synthesis was confirmed by mass spectrometry analysis as sodium adduct ion (m/z=331.1731) and protonated ion (m/z=309.1926) of OD-gal. To compare the antimicrobial activities of OD and newly synthesized OD-gal, MIC values were investigated using E. coli, S. aureus, C. albicans, and A. niger. As a result, it was observed that there was no remarkable difference between MIC values of OD and OD-gal. In addition, to compare the cytotoxicity of OD-gal and OD, HaCaT cells were treated with OD or OD-gal, and then cell viability was quantified using EZ-Cytox assay. In the case of 1.5% OD, the cell viability was 64% at 24 h and 42% at 48 h compared to the control, and cell viability of 1.5% OD-gal-treated cells showed no significant change at 24 h and was 85% at 48 h. Consequently, the cytotoxicity of OD-gal-treated cells was reduced by more than 40% when compared with that of OD-treated cells. Thus, the newly synthesized OD-gal in this study is safer than the existing OD used as a cosmetic additive. In the future, OD-gal will be applicable as a substitute for OD as a less toxic preservative for the cosmetic industry.

• Journal of Ginseng Research
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• v.10 no.2
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• pp.133-140
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• 1986

Ginseng saponin had an effect on the proliferation and viability of cultured murine thymocytes. When the thymocytes were cultured in various concentrations of ginseng saponin, the number of thymocytes increased at $10^{-5}$% ginseng saponin but decreased at $10^{-5}$%. There was little change in the number of thymocytes when cultured in IL 2(Interleukin 2), a factor known for its influence on the proliferation and maturation of thymocytes. When the thymocytes were cultured in various concentrations of IL 2 with $10^{-5}$% ginseng saponin, the number of total cells increased at 1.5% or 3% IL 2 when cultured for 9 hours, or at 6% IL 2 for 12, 24, or 48 hours. But there was little change in the number of viable cells. In vitro, ginseng saponin had an effect on the activity of ADA(Adenosine Deaminase), an enzyme known to affect the production of IL 2. There was a 25% increase in the activity of ADA in the presence of $10^{-5}$% ginseng saponin.

• Journal of Life Science
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• v.31 no.3
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• pp.266-279
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• 2021

The present study examined the cytotoxic effects of a Smilax china L. extract (SCLE) in human cancer (A-549, MCF-7, MDA-MB-231, U87-MG, AGS, MKN-74, and SNU-601) and normal MRC-5 fibroblasts, as well as in mesenchymal stem cells derived from dental tissue (DSC). The 50% inhibitory concentration (IC50) values for SCLE were significantly (p<0.05) lower in the cancer cell lines (A-549, MCF-7, MDA-MB-231, U87-MG, AGS, MKN-74 and SNU-601) than in the MRC-5 and DSC cells. Cell growth was significantly (p<0.05) more inhibited in the cancer cell lines treated with 200 ㎍/ml SCLE than in the normal MRC-5 and DSC, and anoikis-like floating cell morphology was observed in the SCLE-treated cancer cells. The cells detached by SCLE treatment were retrieved daily and assayed for viability and telomerase activity. Cells retrieved at 4 days showed significantly decreased viability and telomerase activity (p<0.05), as well as apoptosis-like abnormal morphology, when compared to cells retrieved in the previous 3 days. The ratio of apoptosis and cells in the G1 phase was significantly (p<0.05) increased in the A-549, AGS, and MCF-7 cancer cells treated with SCLE for 4 days compared to untreated controls. However, after SCLE treatment, cell adhesion was not increased by application of an inhibitor of the associated protein kinase (ROCK) that mainly contributes to the increase in cell attachment. This suggests that the cellular detachment by SCLE is probably controlled by a Rho-independent mechanism(s). These observations indicate that SCLE readily induces anoikis in cancer cells and could serve as a potent agent for cancer chemotherapy.

• Journal of Life Science
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• v.25 no.10
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• pp.1176-1183
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• 2015

Ulmus macrocarpa has been used in Korean medicinal food material to physical disorder or tonic material. The purpose of the present study was to evaluate splenocyte life span expansion effects of Ulmus macrocarpa water extract (UMWE) in general cell culture condition. Splenocytes were handled in the presence of 100 μg/ml UMWE for several different time conditions. Live cells were detected with Hoechst 33,342 dye and cell survival molecules were identified through Western blot. Changes in level of cytokine synthesis were evaluated by ELISA. UMWE showed an effect on increased splenocyte survival. UMWE elevated slightly PI3K phosphorylation and ERK1/2 phosphorylation used at 48 hr and 96 hr. Moreover, Bcl-2 was elevated at 48 hr and 96 hr in UMWE-treated splenocytes. UMWE decreased caspase-3 level at 48 hr and 96 hr. ICAD protein increased at 48 hr culturing time. Hematopoietin IL-2 cytokine was down-regulated, however IL-4 hematopoietin cytokine was up-regulated in UMWE treated cell culture media. Increased IFN-γ levels were verified in the supernatant of UMWE-treated cells in all periods (48 hr and 96 hr). Increased patterns in the production of IL-12 cytokine occurred as compared with control after 48 and 96 hr in UMWE-treated-cell cultures. These results suggested that UMWE can prolong splenocyte life span by controlling various signal factors and cytokines.