• Asian-Australasian Journal of Animal Sciences
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• 제19권1호
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• pp.119-125
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• 2006

The present study including two experiments was designed to determine the effect of media containing different rare earth elements (REE) on proliferation and fatty acids accumulation in 3T3-L1 cell cultures. In Experiment 1, 3T3-L1 preadipocytes in 96-well plates ($1.5{\times}10^4cells/ml$) were cultured with Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS) for 24 h. Then the media were changed to the following 10 different media for 48 h: DMEM containing 10% FBS for the control; the above media containing $5{\mu}M$, $10{\mu}M$ or $15{\mu}M$ of $LaCl_3$, $CeCl_3$ or the mixture of these REE chlorides. The proliferation rate of the cells was measured and compared by a non-isotope method-XTT method. In Experiment 2 the cells in 24-well plates ($1.5{\times}10^4cells/ml$) were cultured in DMEM containing 10% FBS for 7 days until confluent and then were changed to above DMEM containing dexamethasone, methyl-isobutylxanthine and insulin (DMI) for two days. Afterwards the media were changed to the 10 different media with REE supplements as in Experiment 1 and cultured for 6 days. The cells were then harvested for fatty acids analysis by gas chromatography. It was found that supplementation of La (5, 10 and $15{\mu}M$), Ce ($5{\mu}M$ and $15{\mu}M$) and the mixture REE (5, 10 and $15{\mu}M$) stimulated (p<0.05) the proliferation of 3T3-L1 preadipocytes (Experiment 1). In the differentiating 3T3-L1 cells supplementation of La ($5{\mu}M$ and $10{\mu}M$), Ce ($5{\mu}M$) and the mixture REE ($5{\mu}M$ and $15{\mu}M$) decreased (p<0.05) the concentration of monounsaturated fatty acids (MUFA) per $10^5cells$, while the supplementation of La ($5{\mu}M$), Ce ($5{\mu}M$) and the mixture REE ($15{\mu}M$) increased (p<0.05) the ratio of saturated fatty acids (SFA) to MUFA. These results indicate that the supplementation of REE to the media may affect proliferation, differentiation and lipogenesis rates of 3T3-L1 cells. However, the effect may depend upon the level or type of REE applied.

• Asian-Australasian Journal of Animal Sciences
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• 제9권6호
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• pp.701-703
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• 1996

The influence of phenylalanine (Phe) in the medium on protein synthesis of chicken embryo fibroblasts (CEF) was examined. CEF was derived from 9-d-old embryos by trypsin-EDTA digestion. To examine the deficiency of Phe in the medium, CEF was cultured in Dulbecco's modified Eagle's medium (DMEM) with or without Phe. CEF was also cultured in Dulbecco's phosphate buffered saline (PBS ($Ca^{2+}$, $Mg^{2+}$)) with or without $400{\mu}m$ Phe in order to examine the effect of Phe supplementation. All media were supplemented with 10% (v/v) fetal calf serum. After incubation for 6, 30 and 54 h, protein synthesis was measured by the incorporation of L-[2, $6-^{3}H$] Phe into CEF for further 18 h. Protein synthesis of CEF cultured in DMEM was higher than that in PBS ($Ca^{2+}$, $Mg^{2+}$). High specific radioactivity of Phe due to the low concentration of Phe in the medium resulted in the apparent increase in protein synthesis of CEF. Protein synthesis cultured in PBS ($Ca^{2+}$, $Mg^{2+}$) with Phe did not increase during 72 h of cell culture.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2013년도 제44회 동계 정기학술대회 초록집
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• pp.494-494
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• 2013

Atmospheric pressure non-thermal plasma of the needle-typed interaction with aqueous solutions has received increasing attention for their biomedical applications [1]. In this context, surface discharges at bio-solutions were investigated experimentally. We have generated the non-thermal plasma jet bombarding the bio-solution surface by using an Ar gas flow and investigated the emission lines by OES (optical emission spectroscopy) [2]. Moreover, The non-thermal plasma interaction with bio-solutions has received increasing attention for their biomedical applications. So we researched, the OH radical density of various biological solutions in the surface by non-thermal plasma were investigated by Ar gases. The OH radical density of DI water; deionized water, DMEM Dulbecco's modified eagle medium, and PBS; 1x phosphate buffered saline by non-thermal plasma jet. It is noted that the OH radical density of DI water and DMEM are measured to be about $4.33{\times}1016cm-3$ and $2.18{\times}1016cm-3$, respectively, under Ar gas flow 250 sccm (standard cubic centimeter per minute) in this experiment. The OH radical density of buffer solution such as PBS has also been investigated and measured to be value of about $2.18{\times}1016cm-3$ by the ultraviolet optical absorption spectroscopy.

• 한국전자통신학회논문지
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• 제8권10호
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• pp.1595-1600
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• 2013

기존의 기기분석에 있어서의 미비한 점을 보완한 물질 분석 과정에서 주요 성분의 파괴를 최소화하고져 비파괴형 원적외선 도자기 가마를 이용하여 비파괴형 성분물질을 유지할 수 있는 기기 분석 장치를 개발 하였다. 또한 광폭증폭관(PMT) 기기를 사용하여 뽕잎 차에 함유된 항산화 물질의 형태적 효과를 기존의 기기분석기와 원적외선 도자기 기기를 사용한 것과의 차별화 실험을 통해 그 효과를 측정 하였다. 그 결과 특히 과산화수소를 이용한 항산화 효과를 측정한 결과 성분이 파괴되지 않음을 측정하여 그 효과가 인정 되었기에 이를 보고한다.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 2007년도 춘계 학술대회
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• pp.23-24
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• 2007

Purpose: 인삼이 항당뇨 활성을 가진다는 연구가 많은 연구자들에 의해 진행되었고, 이는 인삼의 구성 성분 중 ginsenoside에 기인한다는 보고가 있다. 본 연구는 ginsenoside의 항당뇨 작용기전을 in vitro에서 알아보고자 3T3-L1 지방세포에서 glucose uptake와 췌장 베타세포인 HIT-T15 세포에서 insulin 분비 효과를 확인하였다. 이를 위하여 인삼을 식초로 처리한 긴삼의 70% MeOH 분획으로부터 protopanaxadiol 계인 ginsenoside $Rb_2$, $Rg_3$ 그리고 protopanaxtriol 계인 $Rg_2$를 분리하여 본 실험에 사용하였다. Method: Ginsenoside $Rb_2$, $Rg_2$, $Rg_3$가 지방 세포에서 glucose uptake에 미치는 효과를 확인하기 위하여 3T3-L1 세포를 DMEM (Dulbecco's Modified Eagle's Medium) 배지에서 분화 유도시켰으며 3T3-L1 preadipocyte가 80% 정도 자라면 분화 유도 배지 (5% fetal bovine serum (FBS), 0.5 mM isobutylmethylxanthine (IBMX), 1 mM dexamethasone 그리고 $10{\mu}g/ml$ insulin가 포함된 DMEM)로 4일, $10{\mu}g/ml$ insulin가 포함된 DMEM으로 2일, FBS만 포함된 DMEM으로 2일 배양하여 총 8일 동안 분화를 유도하였다. 분화 유도된 3T3-L1 adipocytes 에 각각 $Rb_2$, $Rg_2$, $Rg_3$를 $20{\mu}M$로 처리하여 16시간 배양하여 low glucose DMEM에서 3시간 배양한 후에 $37^{\circ}C$에서 insulin 10 ng/ml 과 각각 $Rb_2$, $Rg_2$, $Rg_3$가 포함된 Krebs Ringer Hepes buffer(KRP buffer)에서 20분간 배양하였다. 2-deoxy-D-[$^3H$]-glucose를 넣고 10분 후에 차가운 PBS로 반응을 종결시켜 lysis buffer로 cell을 모은 후 scintillation counter를 이용하여 glucose를 측정하였다. Insulin 분비 효과는 HIT-T15 세포와 일차 배양한 흰쥐 소도세포(islets)를 사용하여 확인하였다. HIT-T15 세포는 24 well plate에 well 당 $2{\times}10^5$ 개씩 분주하여 24시간 동안 배양한 후 시료를 처리하였으며 소도 세포는 Sprague-Dawley rat의 췌장에 collagenase가 포함된 Hanks' Balanced Salt Solution(HBSS)을 주입하여 분리하고 islets을 얻었다. 분리한 소도세포를 $1{\sim}2$일 동안 배양하여 $Rb_2$, $Rg_2$, $Rg_3$가 각각 $20{\mu}M$의 농도로 첨가된 insulin 측정용 buffer인 Krebs-Ringer buffer (KRB+0.3% BSA, KRBB)에 $37^{\circ}C$에서 1시간 incubation 시킨 후 배양액으로 분비된 인슐린의 양을 측정하였다. 한편 ginsenoside의 인슐린 분비 촉진 기전을 알아보기 위한 실험에서는 ATP-sensitive $K^+$ channel opener인 diazoxide (0.5 mM)가 ginsenoside에 의해 촉진된 인슐린 분비를 억제하는지 살펴보았다. Result: glucose uptake assay 에서는 $Rg_2$가 가장 크게 glucose uptake를 증가시켰고 $Rb_2$, $Rg_3$는 그 활성이 크지 않았다. 한편 Insulin 분비 효과는 diol계인 $Rg_3$에서 용량 의존적으로 인슐린의 분비를 촉진시켰으며 $20{\mu}M$ 농도에서 대조군과 비교해 1.5배 이상의 분비 촉진 효과를 보였고 triol계인 $Rg_2$ 에서는 이러한 효과가 나타나지 않았다. $Rg_3$의 인슐린 분비 촉진 기전을 0.5 mM 의 diazoxide를 이용하여 확인한 결과 $Rg_3$에 의해 촉진된 인슐린 분비를 감소시켰다. 이로 미루어보아 $Rg_3$의 인슐린 분비 촉진 기전은 ATP-sensitive $K^+$ 채널의 봉쇄에 의한 것임을 확인할 수 있었다.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제32권6호
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• pp.504-510
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• 2010

Purpose: This study was to examine the proliferation and function of the adipose tissue-derived endothelial cells according to different culture medium conditions. Materials and Methods: Adipose tissue-derived CD146 positive endothelial cells were cultured in according to different culture mediums (DMEM culture medium with or without osteogenic inductive agents and EBM-2 culture medium with or without osteogenic inductive agents). The proliferation and function of the adipose tissue-derived endothelial cells was examined in different culture medium conditions. Results: Adipose tissue-derived endothelial cells formed tube-like structures on Matrigel in EBM-2 culture medium with or without osteogenic inductive agents. However, the cells did not form tube-like structures on Matrigel in DMEM medium with or without osteogenic inductive agents. After 24 hours of culture, among the culture medium using EBM-2, the proliferation of the cells were promoted in EBM-2 medium without osteogenic inductive agents than in EBM-2 medium with osteogenic inductive agents. However, 72 hours of culture, the proliferation of the cells were promoted in EBM-2 medium with osteogenic inductive agents than in EBM-2 medium without osteogenic inductive agents. Conclusion: These results suggest that the proliferation and function of the adipose tissue-derived CD146 positive endothelial cells could be maintained in EBM-2 with osteogenic inductive agents.

• Asian-Australasian Journal of Animal Sciences
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• 제28권3호
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• pp.411-419
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• 2015

We previously demonstrated that bovine subcutaneous preadipocytes promote adipogenic gene expression in muscle satellite cells in a co-culture system. Herein we hypothesize that saturated fatty acids would promote adipogenic/lipogenic gene expression, whereas mono- and polyunsaturated fatty acids would have the opposite effect. Bovine semimembranosus satellite cells (BSC) and intramuscular preadipocytes (IPA) were isolated from crossbred steers and cultured with 10% fetal bovine serum (FBS)/Dulbecco's Modified Eagle Medium (DMEM) and 1% antibiotics during the 3-d proliferation period. After proliferation, cells were treated for 3 d with 3% horse serum/DMEM (BSC) or 5% FBS/DMEM (IPA) with antibiotics. Media also contained $10{\mu}g/mL$ insulin and $10{\mu}g/mL$ pioglitazone. Subsequently, differentiating BSC and IPA were cultured in their respective media with $40{\mu}M$ palmitic, stearic, oleic, or linoleic acid for 4 d. Finally, BSC and IPA were single- or co-cultured for an additional 2 h. All fatty acid treatments increased (p = 0.001) carnitine palmitoyltransferase-1 beta ($CPT1{\beta}$) gene expression, but the increase in $CPT1{\beta}$ gene expression was especially pronounced in IPA incubated with palmitic and stearic acid (6- to 17-fold increases). Oleic and linoleic acid decreased (p = 0.001) stearoyl-CoA desaturase (SCD) gene expression over 80% in both BSC and IPA. Conversely, palmitic and stearic acid increased SCD gene expression three fold in co-cultured in IPA, and stearic acid increased $AMPK{\alpha}$ gene expression in single- and co-cultured BSC and IPA. Consistent with our hypothesis, saturated fatty acids, especially stearic acid, promoted adipogenic and lipogenic gene expression, whereas unsaturated fatty acids decreased expression of those genes associated with fatty acid metabolism.

• Journal of Animal Science and Technology
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• 제64권3호
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• pp.481-499
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• 2022

This study aims to determine the effects of D-methionine (D-Met) isomer and the methionine precursor 2-hydroxy-4-methylthiobutanoic acid i (HMBi) supplementation on milk protein synthesis on immortalized bovine mammary epithelial cell (MAC-T). MAC-T cells were seeded using 10-cm dishes and cultured in Dulbecco's modified Eagle's medium/F12 (DMEM/F12) basic medium. The basic medium of DMEM/F12 was replaced with the lactogenic DMEM/ F12 differentiation medium when 90% of MAC-T cells reached confluency. The best dosage at 0.6 mM of D-Met and HMBi and incubation time at 72 h were used uniformly for all treatments. Each treatment was replicated six times wherein treatments were randomly assigned in a 6-well plate. Cell, medium, and total protein were determined using a bicinchoninic acid protein assay kit. Genes, proteomics and metabolomics analyses were also done to determine the mechanism of the milk protein synthesis pathway. Data were analyzed by two-way analysis of variance (ANOVA) with supplement type and plate as fixed effects. The least significant difference test was used to evaluate the differences among treatments. The HMBi treatment group had the highest beta-casein and S6 kinase beta-1 (S6K1) mRNA gene expression levels. HMBi and D-Met treatments have higher gene expressions compared to the control group. In terms of medium protein content, HMBi had a higher medium protein quantity than the control although not significantly different from the D-Met group. HMBi supplementation stimulated the production of eukaryotic translation initiation factor 3 subunit protein essential for protein translation initiation resulting in higher medium protein synthesis in the HMBi group than in the control group. The protein pathway analysis results showed that the D-Met group stimulated fructose-galactose metabolism, glycolysis pathway, phosphoinositide 3 kinase, and pyruvate metabolism. The HMBi group stimulated the pentose phosphate and glycolysis pathways. Metabolite analysis revealed that the D-Met treatment group increased seven metabolites and decreased uridine monophosphate (UMP) production. HMBi supplementation increased the production of three metabolites and decreased UMP and N-acetyl-L-glutamate production. Taken together, D-Met and HMBi supplementation are effective in stimulating milk protein synthesis in MAC-T cells by genes, proteins, and metabolites stimulation linked to milk protein synthesis.

• 한국환경성돌연변이발암원학회:학술대회논문집
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• 한국환경성돌연변이발암원학회 2002년도 Molecular and Cellular Response to Toxic Substances
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• pp.100-105
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• 2002

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2002년도 춘계학술발표대회 발표논문초록집
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• pp.43-43
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• 2002

체세포 핵이식에 의한 산자가 양, 소, 염소, 쥐 및 최근에 돼지에서 보고되었지만, 생산된 배반포의 동결보존에 관한 연구결과는 보고되지 않고 있다. 따라서, 본 연구는 체세포 핵이식란의 활용과 산업화 촉진을 위해서 체세포 핵이식 유래 배반포의 동결성 향상에 배양조건이 미치는 영향을 조사하고자 실시하였다. 공여세포는 60일령의 태아세포를 10% FBS 가 첨가된 DMEM 에서 39℃, 5% CO₂의 incubator에서 배양하여 monolayer confluent 형성을 유도한 후 0.25%trypsin 처리하였다. (중략)