• 한국수정란이식학회지
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• 제18권1호
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• pp.35-41
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• 2003

본 연구에서 배양액 내 첨가된 macromolecule이 돼지 체외수정란의 체외 발달율에 미치는 영향과 배양소적 내 첨가된 FBS가 후기배발달에 미치는 영향에 대하여 알아보았다. 이에 대한 결과를 요약 하면 다음과 같다. 1. NCSU-23을 기본 배양액으로 사용시 BSA (4mg/ml), FBS (10%), PVA (3 mg/ml)가 각각 첨가되었을 경우 배발달에 큰 차이를 보이지 않았다. 2. PZM을 기본 배양액으로 사용시 BSA (4mg/ml)가 첨가하였을 경우 2-세포 수정란 발달율과 배반포 발달율이 가장 높았으며, 이것은 NCSU-23에 비하여 PZM 배양액에는 후기배의 에너지원으로 사용하는 글루코스가 포함되어 있지 않기 때문으로 사료 된다. 본 연구를 통하여 돼지 수정란의 체외배양에 사용되는 배양액은 종류와 조성에 따라 적절한 macromolecule이 첨가되어 초기배와 후기배의 발달을 모두 촉진시킬 수 있어야 함을 알 수 있었다.

• 대한시과학회지
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• 제20권4호
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• pp.531-541
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• 2018

목적 : 항균제 polyhexamethylene Biguanide(PHMB), epigallocatechin gallate(EGCG) 및 PHMB/EGCG 혼합물의 각막상피세포(primary human corneal epithelial cells, HCEpiCs)에 대한 급성독성을 평가하고자 하였다. 방법 : 각막상피세포를 0.00001~0.005% PHMB, 0.001~5% EGCG 및 0.00005% PHMB/0.05% EGCG 혼합물이 각각 포함된 배양액에서 30분, 60분, 120분 및 240분 동안 배양하였다. 배양한 각막상피세포를 고정한 다음 Draq 5로 염색하고 공초점현미경과 ImageXpress $Ultra^{TM}$를 이용하여 세포형태를 관찰하여 세포생존율과 세포자살(apoptosis)을 비교하였다. 결과 : 배양된 각막상피세포는 0.00005% 이하의 PHMB 농도 및 0.05% 이하의 EGCG 농도에서는 세포 독성이 나타나지 않았다. 0.00005% PHMB/0.05% EGCG 혼합물이 포함된 배양액에서 급성 세포독성은 관찰되지 않았으나 240분 배양시킨 경우에는 손상된 각막상피세포 수가 증가하고 생존 세포의 수는 감소하였다. 결론 : 항균 시너지 효과를 갖는다고 보고된 0.00005% PHMB/0.05% EGCG 혼합물의 경우 각막상피세포에 대한 급성독성은 없었으나 만성효과에 대해서는 추가적인 연구가 필요할 것으로 생각된다.

• 한국가축번식학회지
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• 제16권4호
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• pp.347-352
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• 1993

This study was designed to develop the in vitro culture systemof mammalian preimplantation embryos. We proposed mouse fetal fibroblast cells (MFFC) from 14∼15 day mouse fetus. Zygotes from superovulated female ICR mice were cultured 96 hrs in simple defined media (T6) or on the monolayer of MFFC. In addition, to evaluate the action of the co-culture of MFFC, various diluted superoxide dismutase antibody (SOD-Ab) was supplemented into the monolayer of MFFC and zygotes were cultrued in presence or absence of SOD-Ab. The developmental rates of zygotes were significantly increased in co-culture with MFFC compared to the control. The rates of zygotes to the 4-cell stage in media treated with EDTA were higher than those cultured in MFFC but the proportions of morula and blastocyst were not differ between EDTA and MFFC. Interestingly blastocysts in co-culture with MFFC possessed as many as blastomere as those developing in vivo, but blastocysts cultured with EDTA had significantly fewer blastomeres. In addition, the treatment of SOD-Ab suppressed the beneficial effect of MFFC. Therefore, our findings suggest that co-cultrue system using MFFC may have an advantage in the development of mouse zygotes as well as embryonic differentiation.

• Fisheries and Aquatic Sciences
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• 제18권1호
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• pp.81-88
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• 2015

We investigated factors affecting primary cultures of Pacific abalone Haliotis discus hannai ovary-dissociated cells to identify general aspects of their early-phase culture. Ninety-seven cell populations derived from 30 individuals were cultured in different media with varying compositions of medium supplements, and initial attachment, subculture, and survival for ${\geq}10$ weeks were assessed according to medium composition and individual. We also examined the time required for subculture and the rate of cell death according to both culturing period and passage number within 10 weeks. A lack of fetal bovine serum (FBS) and hemolymph significantly inhibited the growth of cultured cells, while we detected no significant effect of medium composition on initial cell attachment. Through data reallocation, with the omission of data from cell populations cultured in FBS-free and hemolymph-free media, we showed that growth inhibition was also affected by individual differences among the abalones used. During the culture, we observed four different types of cell morphology. Moreover, considerable time was required for subculture-18.4 and 19.5 days for first and second subcultures, respectively-and cell death did not occur within 30 days or for passage 0. Our results will provide valuable information for developing universal cell culturing guidelines in abalone species and suggest the feasibility of culturing abalone ovary-dissociated cells.

• 동의생리병리학회지
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• 제17권1호
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• pp.136-139
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• 2003

Cytotoxicity of glucose oxidase(GO) and cardioprotective effect of Salviae Multiorrhizae Radix(SMR) against GO-induced cardiotoxicity were measured for evaluation of cardiotoxicity on cultured mouse pulmonary endotherial cells(PEC) by MTT assay after PEC were cultured for 8 hours at various concentrations of GO. GO was toxic in a time and dose-dependent manner on cultured PEC after PEC were grown for 8 hours in media containing 1～60mU/ml GO. While, cultures were pretreated with 60 μg/ml SMR for 2 hours increased remarkably cell viability. From the above results, it is suggested that GO is toxic on cultured PEC by the decrease of cell viability, and herb medicine such as SMR is very effective in the prevention of vascular toxicity induced by GO.

• 동의생리병리학회지
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• 제17권3호
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• pp.662-665
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• 2003

To examine the cytotoxicity of reactive oxygen species in cultured rat myocardial cells, cytotoxic effect was determined by MTT assay after cultured cells were incubated for 4 hours in the media containing 1～30μM of H₂O₂. And also, the protective effect of Drynariae Rhizoma(DR) was determined in these cultrures. Cell viability was significantly decreased in a dose-dependent manner after exposure of 15 μM H₂O₂ to cultured rat myocardials for 4 hours. In the protective effect of DR, DR prevented the H₂O₂-induced cytotoxicity in these cultures. From these results, it suggests that H₂O₂ has toxic effect in cultured mouse myocardial cells and DR has protective effect on the cytotoxicity induced by H₂O₂.

• 동의생리병리학회지
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• 제17권5호
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• pp.1305-1308
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• 2003

To evaluate the neurotoxicity of reactive oxygen species (ROS) in cultured cultured spinal dorsal root(DRG) neurons derived from neonatal mouse, Cytotoxicity was measured by MTS assay after cultured cells were grown for 3 hours in the media containing 1～60 μM hydrogen peroxide (H₂O₂). In addition the neuroprotective effect of Salviae Miltiorrhzae Radix (SMR) was measured in these cultrures. Cell viability was positively decreased in a dose- and time-dependent manner after exposure of cultured mouse DRG neurons to 30 tt M H202 for 3 hours. In the neuroprotective effect of SMR on H₂O₂-mediated toxicity, SMR prevented the H₂O₂-induced neurotoxicity in these cultures. From these results. it suggests that H₂0₂ is toxic in cultured mouse spinal motor neurons and selective herb extract such as Uncariae Ramulus Cum Uncis is effective in prevetion of the neurotoxicity induced by H₂O₂.

• Journal of Microbiology and Biotechnology
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• 제12권5호
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• pp.740-745
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• 2002

The current study was conducted to identify the differences in the rheological properties of Chlorella sp. powder cultured in a fermentor and in a pond-like environment. Cells. cultured in the same media were harvested and spray dried. The biomass yield from the fermentor culture was 4.7% (dry basis), while that from the pond was 4.3% (dry basis). Measurements of the loose bulk density, tapping test, Hausner's ratio, and compressibility test all revealed differences between the rheological properties of the Chlorella sp. from the two cultivation systems. Although both the fermentor and pond cultured Chlorella sp. showed the same angle of repose, the mean size of the cells was 2.26 $\mu\textrm{m}$ and 2.89 $\mu\textrm{m}$, respectively. The weight of the Chlorella sp. tablets cultured in the fermentor and pond was 0.663 g/tablet and 0.593 g/tablet, respectively, while the friability of the tablets was 21% and 41%, respectively. Observation by Transmission Electron Microscope (TEM) showed that the cell wall of the Chlorella sp. cultured in the fermentor was thinner and more spherical than that cultured in the pond, thereby providing the main characteristic rheological properties of the powder.

• 동의생리병리학회지
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• 제16권3호
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• pp.553-556
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• 2002

To study the effects of Scorpio on oxygen free radical-mediated damage by xanthine oxidase/hypoxanthine (XO/HX) on cultured spinal sensory neurons, in vitro assays such as MTT assay were used in cultured spinal sensory neurons derived from mice. Spinal sensory neurons were cultured in media containing various concentrations of XO/HX for 6 hours, after which the neurotoxic effect of XO/HX was measured by in vitro assay. The protective effect of the herb extract, Scorpio water extract against XO/HX-induced neurotoxicity was also examined. The results are as follows : In MTT assay, XO/HX significantly decreased the cell viability of cultured mouse spinal sensory neurons according to exposure concentration and time in these cultures. The effect of Scorpio water extract on XO/HX-induced neurotoxicity showed a quantitative increase in neurdfilament. These results suggest that XO/HX has a neurotoxic effect on cultured spinal sensory neurons from mice and that the herb extract, Scorpio water extract, was very effective in protecting XO/HX-induced neurotoxicity.

• 동의생리병리학회지
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• 제17권2호
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• pp.499-502
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• 2003

Neurotoxicity of reactive oxygen species(ROS) and neuroprotective effect of Aconiti Radix(AR) against ROS-induced cytotoxicity were determined on cultured mouse cerebral neurons by MTT assay after cerebral neurons were cultured for 5 hours in various concentrations of GO. GO was toxic in a dose-dependent manner on cultured cerebral neurons after cerebral neurons were incubated for 5 hours in media containing 5～40mU/ml GO. While, cultures were pretreated with 180 μg/ml AR for 2 hours increased remarkably cell viability. From these results, it is suggested that GO has toxic effect on cultured mouse cerebral neurons by the decrease of cell viability. And also, herb extract such as AKR is very effective in the protection pf neurotoxicity induced by GO.