• 제목/요약/키워드: Serum-free Medium

검색결과 192건 처리시간 0.029초

TGF-$\beta$1와 IGF-I이 소 난포란의 체외성숙 및 체외수정란의 배양에 미치는 영향 (Effect of TGF-$\beta$1 and IGF-I on Bovine In Vitro Maturation and Embryo Culture)

  • 서태광
    • 한국가축번식학회지
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    • 제20권2호
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    • pp.111-117
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    • 1996
  • 본 연구는 혈청첨가 또는 무첨가에 따른 소 난포란의 체외성숙에 있어서 참가된 TGF-$\beta$1과 IGF-I이 그후의 수정 및 발생에 미치는 영향과, 이들 growth factor의 농도에 따른 8세포기 소 체외수정란의 발달에 미치는 영향을 조사하고자 실시하였다. 도축장에서 얻어진 난소로부터 채취된 난포란을 20% FBS가 첨가 또는 첨가되지 않은 TCM-199에 TGF-$\beta$1, IGF-I 또는 TGF-$\beta$1+IGF-I을 각각 10ng/ml 첨가하여 38.5$^{\circ}C$에서 24시간 배양하여 체외성숙을 유기하였다. 성숙된 난자를 1$\times$106/ml 정자농도로 수정후 24시간에 glucoserk 첨가되지 않은 CZB 배양액으로 옮겨 48시간 배양한 다음, TCM-199+20%FBS에서 96시간 추가배양하였다. 본 연구에서 혈청이 첨가된 난포란의 체외성숙배양액에 첨가된 growth factor들은 수정후의 배분할 및 배발생에 영향을 미치지 않았다. 혈청이 첨가되지 않은 경우에서는 TGF-$\beta$1의 첨가는 배분할 및 배발생율을 향상시켰다(P<0.05). 한편 TCM-199+20%FBS에 5, 10ng/ml의 TGF-$\beta$1 및 5, 10, 50, 100ng/ml의 IGF-I을 각각 첨가후 8세포기 체외수정란을 배양한 결과, 10ng/ml TGF-$\beta$1의 첨가는 배반포기로의 발생율을 향상시켰다(P<0.05). 결론적으로, 혈청이 포함되지 않은 소 난포란의 체외성숙 배양액, 또는 수정란의 체외배양액에 10ng/ml TGF-1의 첨가는 배반포기로의 발생율을 향상시킨다.

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Differential Regulation of Obesity by Swim Training in Female Sham-operated and Ovariectomized Mice

  • Jeong, Sun-Hyo;Yoon, Mi-Chung
    • 대한의생명과학회지
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    • 제17권1호
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    • pp.13-20
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    • 2011
  • The peroxisome proliferator-activated receptor ${\alpha}$ ($PPAR{\alpha}$) is a nuclear transcription factor that plays a central role in lipid and lipoprotein metabolism. To investigate whether swim training improves obesity and lipid metabolism through $PPAR{\alpha}$ activation in female sham-operated (Sham) and ovariectomized (OVX) mice, we measured body weight, visceral adipose tissue mass, serum free fatty acid at 6 weeks as well as the expression of hepatic $PPAR{\alpha}$ target genes involved in fatty acid oxidation. Swim-trained mice had decreased body weight, visceral adipose tissue mass and serum free fatty acid levels compared to high fat diet fed control mice in both female Sham and OVX mice. These reductions were more prominent in OVX than in Sham mice. Swim training significantly increased hepatic mRNA levels of $PPAR{\alpha}$ target genes responsible for mitochondrial fatty acid ${\beta}$-oxidation, such as carnitine palmitoyltransgerase-1 (CPT-1), very long chain acyl-CoA dehydrogenase (VLCAD), and medium chain acyl-CoA dehydrogenase (MCAD) in OVX mice. However, swim trained female Sham mice did not increase hepatic mRNA levels of $PPAR{\alpha}$ target genes responsible for mitochondrial fatty acid ${\beta}$-oxidation compared to Sham control mice. These results indicate that swim training differentially regulates body weight and adipose tissue mass between OVX and Sham mice, at least in part due to differences in liver $PPAR{\alpha}$ activation.

신장 조직의 브라디키닌 결합부위 발현 (Expression of Bradykinin Binding Sites within the Mammalian Kidney Tissues)

  • 정성현;정지창
    • 약학회지
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    • 제38권5호
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    • pp.602-607
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    • 1994
  • Based upon the previous experiments showing that kidney and lung tissues of rat had relatively abundant bradykinin binding sites, we tried to characterize and determine the densities of the bradykinin binding sites in the rabbit kidney tissue and proximal tubular cells under different growing conditions. Among the kidney tissue renal medulla segments showed the highest bradykinin binding sites. To determine which growth factors are to add in the serum free culture medium to express selectively the bradykinin binding sites in the rabbit kidney proximal tubular cells, we tried so called hormone-deletion approach and in here insulin, hydrocortisone, transferrin, triiodothyronine and prostaglandin $E_1$ are examined. By performing receptor binding assay and determination of protein concentrations, we may conclude that the most required hormones in the expression for bradykinin binding sites are insulin and transferrin, and fetal bovine serum is shown to be less effective in this regard.

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Conjugated Linoleic Acid에 의한 대장암 세포 증식 억제 기전 연구 (Study of the Mechanism for the Growth Inhibitory Effects of Conjugated Linoleic Acid on Caco-2 Colon Cancer Cells)

  • 김은지;오윤신;이현숙;박현서;윤정한
    • Journal of Nutrition and Health
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    • 제36권3호
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    • pp.270-279
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    • 2003
  • Conjugated linoleic acid (CLA) is a group of positional and geometric isomers of linoleic acid (LA) and exhibits anticarcinogenic activity in a variety of animal models. We have previously observed that CLA inhibited the growth of Caco-2 cells, a human colon adenocarcinoma cell line. The present study was performed to determine whether the growth inhibitory effect of CLA is related to change in secretion of IGF- II and/or IGF-binding proteins (IGFBPs) that have been shown to regulate Caco-2 cell proliferation by an autocrine mechanism. Cells were incubated in serum-free medium with various concentrations of CLA or linoleic acid (LA). Immunoblot analysis of 24-hours, serum-free, conditioned medium using a monoclonal anti-IGF-IIantibody revealed that Caco-2 cells secreted both mature 6,500 Mr and higher Mr forms of pro IGF-II. The levels of pro IGF-II and mature IGF-IIwere decreased by 43 $\pm$ 2% and 53 $\pm$ 6%, respectively by treatment with 50 $\mu$ M CLA. LA slightly increased pro IGF- II levels. Results from Northern blot analysis showed that CLA decreased IGF-II mRNA levels at 50 $\mu$ M concentration suggesting that CLA regulation of IGF-II protein expression occurs partly at the transcriptional level. Ligand blot analysis of conditioned media using 1251-IGF-II revealed that CLA slightly decreased IGFBP-2 levels and increased IGFBP-4 levels. We confirmed our previous results that CLA inhibited cell growth in a dose-dependent manner but LA slightly increased cell growth. Exogenous IGF-II mitigated the growth inhibitory effect of CLA. These results indicate that the growth inhibitory effect of CLA may be at least in part mediated by decreasing IGF-II and IGFBP-2 secretion and increasing IGFBP-4 secretion in Caco-2 cells.

교반형 막 반응기를 이용한 재조합 인간 세포의 무혈청 배지에 의한 $\gamma$-Interferon의 생산 (Economic Production of $\gamma$-Interferon from Recombinant Human Cells in Serum Free Medium by a Moving Aeration Membrane Bioreactor)

  • 박영식;김현규;임서규;박경유;이현용
    • 한국미생물·생명공학회지
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    • 제22권4호
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    • pp.389-394
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    • 1994
  • 8 X 10$^{6}$(viable cells/ml) of maximum cell density and 9000(IU/ml) of $\gamma$-IFN production were obtained at 55(ml/hr) of a perfusion rate by cultivating HSF cells using a moving membrane aeration bioreactor. This system proves to be an efficient culture process by maintaning 90% of viable cells during the whole cultivation periods. The metabolic molar quotient of glucose to lactate was 0.81 for overall ranges of glucose consumed while the evolution of ammonia was not linearly related to the consumption of glutamine. Low molar conversion ratio was observed in low consumptions of glutamine and high molar conversion ratio in high comsumptions. It also shows that the glutamolysis plays important role in the steady state conditions by evolving larger quantities of ammonia than lactate. At the above of 50 rpm, which is the optimum agitation speed for this bioreactor, the cell growth was severely affected while the IFN production was less decrea- sed, maintaing 1.5 X 10$^{-3}$(IU/cell/day) specific IFN production rate. The cumulatvie $\gamma$-IFN production was 7.2 X 10$^{8}$(IU) for 70 days of the cultivation, which yields 1 X 10$^{7}$ (IU/day) of IFN production rate. Therefore, a commercial production of $\gamma$-IFN by this culture process can be achievable by maintaining the above IFN productivity in a scaled-up culture system.

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Bovine Oocytes Can Be Penetrated in Modified Tris-buffered Medium

  • Park, Kwang-Wook;Niwa, Koji
    • Asian-Australasian Journal of Animal Sciences
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    • 제22권4호
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    • pp.500-506
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    • 2009
  • A modified Tris-buffered medium (mTBM) has been widely used as an insemination medium for porcine in vitro fertilization (IVF). We examined whether mTBM could be used for bovine IVF. Bovine cumulus-oocyte complexes (COCs) were cultured in a serum-free medium containing 30 ng/ml EGF for 22 h. After culture, COCs were inseminated with spermatozoa for 12 h in mTBM containing 5 mM caffeine and 10 g/ml heparin. The penetration of oocytes increased significantly (p<0.05) as the sperm concentration increased from 0.1 (30%) to 1-10 $(87-100%){\times}10^6$ cells/ml. This was significantly different from values obtained at 1 (87%) and 10 $(100%){\times}10^6$ cells/ml. However, when COCs were inseminated with spermatozoa from different bulls, the proportions (62-100%) of oocytes penetrated varied according to the bull. The proportion (18%) of oocytes penetrated was significantly (p<0.05) lower in a fertilization medium without caffeine and heparin but increased with the addition of caffeine and/or heparin to the medium, and the proportion (93-96%) of oocytes penetrated increased significantly (p<0.05) when the medium was supplemented with heparin and caffeine. In this medium, sperm penetration was first observed at 3 h after insemination. Irrespective of the presence of glucose in the fertilization medium, the proportion (93-97%) of oocytes penetrated and the proportion (83-84%) of embryos at the ${\geq}2$-cell stage cultured in a chemically defined medium were not significantly different. However, the proportion of embryos developing to the blastocyst stage was significantly (p<0.05) higher in the presence (11%) of glucose in the fertilization medium than in its absence (2%). In conclusion, the present study demonstrated that bovine oocytes penetrated in vitro in mTBM can develop to the blastocyst stage and mTBM may be used for the in vitro production of bovine embryos.

BcI-2 Over-expression Reduced the Serum Dependency and Improved the Nutrient Metabolism in a NS0 Cells Culture

  • Tey Beng Ti;Al-Rubeai Mohamed
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제10권3호
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    • pp.254-261
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    • 2005
  • The over-expression of Bcl-2 has greatly improved the culture period, specific growth rate, and maximum viable cell density of NS0 cells culture under low serum condition. Further analysis of these data suggests that a saturation model of the Monod type can be used to represent the relationships of specific growth rate and initial serum concentration. The ${\mu}_{max}$ and $K_s$ for the Bcl-2 cell line is $0.927day^{-1}\;and\;0.947\%(v/v)$ respectively, which are $21\%$ greate and $7\%$ lower respectively than its control counterpart. Study on the amino acid supplementation revealed that Bcl-2 cell lines possess greater improvement in the specific growth rate and maximum viable cell density compared to the control cell lines. A further increase in the amino acid supplementation has resulted a $17\%$ decrease in specific growth rate and no improvement in maximum viable cell density in the control culture. However, the Bcl-2 cell line exhibited a better growth characteristic in this culture condition compared to that of control cell lines. The higher specific growth rate and maximum viable cell density of the Bcl-2 cell line in medium fortified with serum and MEM EM suggested a more efficient nutrient metabolism compared to that in the control cell line. The low serum and amino acid utilisation rate and the higher cell yield may prove to be important in the development of serum/protein free culture.

중쇄(中鎖)와 장쇄지방(長鎖脂肪) 식이(食餌)가 Cholesterol급여(給與) 흰쥐의 혈청지질(血淸脂質)에 미치는 영향(影響) (Effect of Medium and Long Chain Triglyceride Diet on the Serum Lipids of Rats Fed with Cholesterol)

  • 이은숙;조정순
    • 한국응용과학기술학회지
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    • 제4권1호
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    • pp.91-97
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    • 1987
  • Because of its more complete hydrolysis and rapid absorption, MCT is expected its usefulness in the diets of patients with malabsorption syndrom. Also, several authors reported that serum cholesterol level was lower after MCT feedings. In this study, rate of each experimental group were fed for 4weeks with 20% MCT, 20% corn oil, mixed diet of 10% MCT and 10% corn oil, mixed diet of 17% corn oil and 3% shortening. After experimental diet, it was measured to growth rate and serum cholesterol, triglyceride and phospholipid level. The results were as follows; 1. The body weight gain was the lowest in MCT group and others were lower than control group. 2. Serum total cholesterol level was the lowest in corn oil group and others were significantly lower than control group. Free cholesterol level was lower in all experimental groups than control group too. 3. Serum HDL-cholesterol level was significantly higher in all experimental groups than control group. 4. The ratio of VLDL, LDL-Cholesterol to HDL-Cholesterol was significantly lower in all experimental groups than control group. 5. Serum triglyceride level was higher in all experimental groups than control group. Serum phospholipid level was significantly in only mixed diet group of corn oil and shortening than control group. 6. The ratio of total cholesterol to phospholipid was significantly lower in corn oil diet group than control group.

Ginsenoside Rgi is an Anti-apoptotic Agent

  • Zhang, Jun-Tian;Li, Jun-Qing
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 1998년도 Advances in Ginseng Research - Proceedings of the 7th International Symposium on Ginseng -
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    • pp.12-20
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    • 1998
  • Primary neuronal culture was studied for observing effect of ginsenoside Rgl (Rgl) on serum-free medium induced apoptosis. Results showed that Rgl at concentration of 1 umol$.$ L-1 and 10 umol$.$L-1 could inhibit apoptosis, decrease intracellular calcium concentration in cultured cortical neurons, enhance SOD activity in both aged rat cortex and cultured cortical neurons, scavenge cytotoxic oxygen free radicals, decrease NO content and NOS activity in aged rat cortex and cultured cortical neurons, increase bel-2 gene expression in rat brain. These results provided new data for elucidating the anti-aging effect of Rgi. Rgl is considered to be a useful drug for treatment of Alzheimer's disease and brain aging.

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단일지방산 첨가에 의한 간세포의 지질조성과 지방산조성에 미치는 영향 (Effects of Stearic, Oleic and Elaidic Acid on Cellular Lipids and Their Fatty Acid Composition in Hep-$G_2$ Cells)

  • 김대진;조병희
    • 한국식품영양과학회지
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    • 제25권3호
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    • pp.399-405
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    • 1996
  • The effects of stearic(18 : 0, SA), oleic(18 : 1 cis, OA) and elaidic acid(18 : 1 trans, EA) on the cell growth, contents of cellular lipids, and the fatty acid composition of cellular and medium lipids in Hep-G$_2$cells were evaluated. The cells were incubated in serum-free medium containing 25, 50, 100 and 200$\mu$M of a fatty acid combined with albumin for 2 days. The fatty acid concentration up to 100$\mu$M showed the normal growth, but the cell growth decreased in the presence of 200$\mu$M fatty acid. The treatment of cells with 100$\mu$M of a fatty acid for two days significantly(p<0.05) increased the cellular triglyceride(TG) content in all fatty acid groups compared to control, but TG contents was not significantly different among all treatment group, but total cholesterol(TC) was the highest level in EA group. The level of free cholesterol(FC) and cholesteryl ester(CE) was similar to those of TC in all fatty acid treated group. The cellular phospholipid(PL) contents were similar between the control and all fatty acid groups. The treatment of cells with SA has no notable effects on the fatty acid composition of TG, CE and PL. The OA treatment caused significant increases in CE(51.2%) and PL(29.8%), but not in TG. The EA treatment resulted in 10.1, 10.7 and 7.8% of $C_{18:1\;trans}$ content in cellular TG, CE, and PL. The TG, CE and PL of medium were relatively similar between SA and OA groups. In EA treated group, TG, CE and PL of medium contained 17.0%, 0.7% and 5.6% of $C_{18:1\;trans}$, respectively.

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