• Title/Summary/Keyword: Optimal culture

Search Result 1,892, Processing Time 0.03 seconds

The optimal conditions to improve retrovirus-mediated transduction efficiency to NIH 3T3 cells (레트로바이러스(retrovirus)의 NIH 3T3 세포로의 유전자 전달효율을 증가시키기 위한 적절한 조건들)

  • Lee, Jun Ah;Lee, Kang-Min;Lee, Hyun Jae;Lee, Yun Jeong;Kim, Dong Ho;Lim, Jung Sub;Park, Kyung-Duk
    • Clinical and Experimental Pediatrics
    • /
    • v.50 no.10
    • /
    • pp.1011-1017
    • /
    • 2007
  • Purpose : We tried to assess the optimal conditions to improve low transduction efficiency and their effect on target cells. Methods : Cultured NIH 3T3 cells were incubated with retroviral vectors bearing an enhanced green fluorescent protein (eGFP) gene. We varied the ratio of viral vectors to target cells (1:1-1:8) and the number of transfections (${\times}1$, ${\times}2$), and compared transduction efficiencies. Also, the effects of polybrene on transduction efficiency and viability of target cells were assessed. Transduction of the eGFP gene was evaluated by observing NIH 3T3 cells under a fluorescence microscope and efficiencies were measured by the percentage of eGFP positive cells using FACscan. Results : As the ratio of retroviral vectors to target cells increased, transduction efficiency was greatly improved, from 7% (1:1) to 38% (1:4). However, transduction efficiency did not increase any more when the ratio increased from 1:4 to 1:8. Cells transfected twice showed higher transduction efficiencies than cells transfected once, at a ratio of 1:8. The eGFP gene transduced to NIH 3T3 cells sustained its expression during repeated passages. However, after the third passage (day 9), the percentage of eGFP positive cells began to decline. The degree of this decline in eGFP expression was lower in cells transfected twice than in cells transfected once (P<0.05). The addition of polybrene did not have any toxic effect on NIH 3T3 cells and greatly increased transduction efficiency (P=0.007). In addition to vector component, transduction efficiency was very sensitive to culture confluence. Cells cultured and transfected in 24-well plate showed higher transduction efficiency, although cells cultured in 6- well plate proliferated more (P=0.024). Conclusion : Our data could be used as a basis for retrovirus-based gene therapy. Further study will follow using human cells as target cells.

Effects of Manipulation Conditions on Development of Nuclear Transplant Bovine Embryos Derived from In Vitro Matured Oocytes (미세조작조건이 소 핵이식배의 발달에 미치는 영향)

  • 최상용;노규진;공일근;송상현;조성근;박준규;이효종;박충생
    • Korean Journal of Animal Reproduction
    • /
    • v.21 no.3
    • /
    • pp.293-302
    • /
    • 1997
  • Follicular oocytes of Grade I and II were collected from 2~6 mm ovarian follicles and matured in vitro (IVM) for 24 hrs in TCM-199 su, pp.emented with 35$\mu\textrm{g}$/ml FSH, 10$\mu\textrm{g}$/ml LH, and 1$\mu\textrm{g}$/ml estradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. They were fretilized in vitro (IVF) by epididymal spermatozoa capacitated with heparin for 12 hrs. The zygotes were then co-cultured in vitro with bovine oviducted epithelial cells (BOEC) for 7 to 9 days. The optimal time for IVM, the successful enucleation of IVM oocytes by micromanipulation at different oocyte ages after IVM, and the ideal culture system for IVM for effective IVF and in vitro development of IVM-IVF embryos was examined for in vitro production of nuclear recipient oocytes and nuclear donor embryos. To improve the efficiency of nuclear transplantation (NT) of IVF embryo into IVM follicular oocytes, this study evaluated the optimal electric condition and oocytes age for activation of IVM oocytes and in vitro development of NT embryos. In vitro development of NT embryos with preactivation or non-preactivation in enucleation oocytes, cell number of IVN-IVF embryos, and NT embryos wre also examined. The results obtained were as follows; 1. The most suitable enucleation time was at 24 hpm (83.3%) rather than that of 28 hpm(69.6%) and 32 hpm(50.0%). 2. There was no difference among the fusion rates of NT embryos at the voltages of 0.75, 1.0 and 1.5 kV/cm, but the in vitro development rates to morule and blastocyst were significantly (P<0.05) higher at the voltage of 0.75(12.5%) and 1.0kV/cm (12.6%) compared to 1.5kV/cm(0%). 3. No significant difference in activation rates were seen in NT embryos stimulated for 30, 60 and 120 $\mu$sec (71.7, 85.2 and 71.9%, respectively), but the in vitro development rates to morulae and blastocyst were significantly (P<0.05) higher in the oocytes stimulated for 30 $\mu$sec (11.6%) and 60 $\mu$sec(10.7%) than 120 $\mu$sec(0.0%). 4. The fusion rates (71.0 and 87.3%) and the in vitro development rates (9.1 and 12.7%) to morula and blastocyst were seen in the NT embryos stimulated at 28 and 32 hpm under the condition of 1.0 kV/ml, 60 $\mu$sec. However, at 24 hpm the fusion rates were 64.8% and the in vitro development to morula and blastocyst were not seen. 5. The fusion rates between the 8~12, 13~17 and 18~22-cell stage of IVM-IVF embryos were not significantly different. The in vitro development rates of the fused embryos to morula and blastocyst which were received from a blastomere of 8~12, 13~17 and 18~22-cell stages of IVM-IVF embryos were 14.9, 8.3 and 6.5%, respectively. 6. The in vitro development rate of the enucleated recipient oocytes with preactivation (24.2%) to morula and blastocyst was significantly (P<0.05) higher than that of non-preactivation (12.8%). 7. The cell numbers of NT blastocyst and IVM-IVF blastocyst cultured during 7~9 days were 63$\pm$11 and 119$\pm$23, and then their the mean cell cycle number were 5.98 and 6.89, respectively.

  • PDF

Evaluation of Pea (Pisum sativum L.) Genetic Resources for Sprout and Analysis of Optimum Culture Condition for Pea Sprout (완두나물 재배에 적합한 유용자원 선발과 완두나물 적정 재배조건 설정)

  • Koo, Sung-Cheol;Kang, Beom-Kyu;Kim, Hyun-Tae;Seo, Jeong-Hyun;Jeong, Kwang-Ho;Yun, Hong-Tae;Oh, In-Seok;Choi, Man-Soo
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.60 no.4
    • /
    • pp.484-490
    • /
    • 2015
  • Thirty-eight Pea (Pisum sativum L.) genotypes were screened to identify varieties to be suitable for sprout. Based on seed yield and sprout qualities such as whole length and sprout yield, five genotypes (PI269803, PI343278, PI343283, PI343300 and PI 343307) were primarily selected as candidates for pea sprouts. In order to determine optimal cultivation condition for pea sprouting, growth characteristics were investigated according to the change of germination temperature and days for sprouting. Whole length and hypocotyl length were observed to increase as a time dependent manner at each tested temperature (20, 23, and $25^{\circ}C$). However, whole length, hypocotyl length, and sprout yield were highly increased at $23^{\circ}C$ compared to 20 and $25^{\circ}C$. Especially, PI269803 and PI343300 showed higher sprout yield than the others. In addition, the effect of the change of germination temperature on antioxidant properties was estimated by measuring total phenolic content (TPC) and free radical scavenging activity (DPPH and ABST activity). TPC and DPPH/ABST activities of PI269803 and PI343300 were higher at $23^{\circ}C$ than at 20 and $25^{\circ}C$, while antioxidant properties of PI343278 and PI343283 were decreased in a temperature-dependent manner. The results show a high degree of correlation between TPC and antioxidant activities and suggest that the temperature change for pea sprouting could be responsible for antioxidant properties. Taken together, these results provide optimal cultivation conditions for pea sprouting and suggest that PI269803 and PI343300 with high sprout yield and antioxidant properties could be used for pea sprouts.

Comparison in Serum Constituents of Cultured Marine Fishes in Early Summer Season (주요 양식어류의 하절기 혈액성분 비교)

  • 전중균;김형배
    • Journal of Aquaculture
    • /
    • v.11 no.4
    • /
    • pp.547-556
    • /
    • 1998
  • This study was performed to obtain the basic data on the serum constituents of several marine fish spesies commonly cultured in Korea. Blood samples taken from six species of fish were analyzed for various components of serum, total protein (TP), albumin (ALB), triglyceride (TRIG), cholesterol (CHOL), glucose (GLC), lipase (LIPA), amylase (AMYL), aspartate transaminoferase (AST), sodium (Na), potassium (K), chloride (CI) and phosphorus (PHOS). The fish used were coho salmon (Oncorhynchus kisutch), rock fish (Sebastes schlegeli), sea bass (Lateolabrax japonicus), olive flounder (Paralichthys olivaceus), rock fish (Sebastes schlegeli), sea bass (Lateolabrax japonicus), olive flounder (Paralichthys olivaceus), parrot fish (Oplegnathus fasciatus) and jack mackerel (Trachurus jaonicus) reared at the Chungmu Experimental Fish Culture Station of KORDI when the water tempetature was ca. 16.5$^{\circ}C$. There were significant differences in TRIG, CHOL, LIPA and AMYZ among the species analyzed. TRIG concentratin were ranged 178~180mg/dl in jack mackeerel and rock fish, 126~159 mg/dl in olive flounder and sea bass, and 102~114 mg/dl in coho salmon and parrot fish, respectively. Jack mackerel showed the highest levels in CHOL (255mg/dl) and GLC(138mg/dl) among species. LIPA levels were recorded 256 U/dl in coho salmon, 41~42 U/dl in parrot fish and rock fisk, and 5~11 U/dl jack mackerel and sea bass, respectively. AMYL activity of coho salmon was measured as 2, 665 U/dl, and that of jack mackerel was 1,210 U/dl while sea bass showed 60 U/dl and parrot fish, olive flounder and rock fish had at most 5 U/dl. On the other hand, there was no significant difference in the concentration of Na and CI. Na and K were proved that they were negatively correlated in all the species. Generally, among blood components, PHOS and CHOL levels were different depending on environmental temperature of each fish species, especially in olive flounder. Rock fish and parrot fish showed high blood concentration of those components during low temperature period while olive flounder and jack mackerel reached high level during their optimal environmental temperature period. The electrolyte concentration and LIPA activity were high during low water temperature period, in general, but TP and ALB concentrations were high during optimal temperature period. The concentrations of TRIG, CHOL and GLC, those which were used as energy sourses, were different among species by season.

  • PDF

Isolation and Characteristics of a Phenol-degrading Bacterium, Rhodococcus pyridinovorans P21 (페놀분해세균 Rhodococcus pyridinovorans P21의 분리 및 페놀분해 특성)

  • Cho, Kwang-Sik;Lee, Sang-Mee;Shin, Myung-Jae;Park, Soo-Yun;Lee, Ye-Ram;Jang, Eun-Young;Son, Hong-Joo
    • Journal of Life Science
    • /
    • v.24 no.9
    • /
    • pp.988-994
    • /
    • 2014
  • The effluents of chemical and petroleum industries often contain non-biodegradable aromatic compounds, with phenol being one of the major organic pollutants present among a wide variety of highly toxic organic chemicals. Phenol is toxic upon ingestion, contact, or inhalation, and it is lethal to fish even at concentrations as low as 0.005 ppm. Phenol biodegradation has been studied in detail using bacterial strains. However, these microorganisms suffer from substrate inhibition at high concentrations of phenol, whereby growth is inhibited. A phenol-degrading bacterium, P21, was isolated from oil-contaminated soil. The phenotypic characteristics and a phylogenetic analysis indicated the close relationship of strain P21 to Rhodococcus pyridinovorans. Phenol biodegradation by strain P21 was studied under shaking condition. The optimal conditions for phenol biodegradation by strain P21 were 0.09% $KNO_3$, 0.1% $K_2HPO_4$, 0.3% $NaH_2PO_4$, 0.015% $MgSO_4{\cdot}7H_2O$, 0.001% $FeSO_4{\cdot}7H_2O$, initial pH 9, and $20-30^{\circ}C$, respectively. When 1,000 ppm of phenol was added to the optimal medium, the strain P21 completely degraded it within two days. Rhodococcus pyridinovorans P21 could grow in up to 1,500 ppm of phenol as the sole carbon source in a batch culture, but it could not grow in a medium containing above 2,000 ppm. Moreover, strain P21 could utilize toxic compounds, such as toluene, xylene, and hexane, as a sole carbon source. However, no growth was detected on chloroform.

Optimization of Medium for the Carotenoid Production by Rhodobacter sphaeroides PS-24 Using Response Surface Methodology (반응 표면 분석법을 사용한 Rhodobacter sphaeroides PS-24 유래 carotenoid 생산 배지 최적화)

  • Bong, Ki-Moon;Kim, Kong-Min;Seo, Min-Kyoung;Han, Ji-Hee;Park, In-Chul;Lee, Chul-Won;Kim, Pyoung-Il
    • Korean Journal of Organic Agriculture
    • /
    • v.25 no.1
    • /
    • pp.135-148
    • /
    • 2017
  • Response Surface Methodology (RSM), which is combining with Plackett-Burman design and Box-Behnken experimental design, was applied to optimize the ratios of the nutrient components for carotenoid production by Rhodobacter sphaeroides PS-24 in liquid state fermentation. Nine nutrient ingredients containing yeast extract, sodium acetate, NaCl, $K_2HPO_4$, $MgSO_4$, mono-sodium glutamate, $Na_2CO_3$, $NH_4Cl$ and $CaCl_2$ were finally selected for optimizing the medium composition based on their statistical significance and positive effects on carotenoid yield. Box-Behnken design was employed for further optimization of the selected nutrient components in order to increase carotenoid production. Based on the Box-Behnken assay data, the secondary order coefficient model was set up to investigate the relationship between the carotenoid productivity and nutrient ingredients. The important factors having influence on optimal medium constituents for carotenoid production by Rhodobacter sphaeroides PS-24 were determined as follows: yeast extract 1.23 g, sodium acetate 1 g, $NH_4Cl$ 1.75 g, NaCl 2.5 g, $K_2HPO_4$ 2 g, $MgSO_4$ 1.0 g, mono-sodium glutamate 7.5 g, $Na_2CO_3$ 3.71 g, $NH_4Cl$ 3.5g, $CaCl_2$ 0.01 g, per liter. Maximum carotenoid yield of 18.11 mg/L was measured by confirmatory experiment in liquid culture using 500 L fermenter.

Preparation and Characterization of Enzymatic Oyster Hydrolysates-added Yogurt (굴 효소 가수분해물 첨가 요구르트의 제조 및 특성)

  • Chung, In-Kwon;Kim, Hye-Suk;Kang, Kyung-Tae;Choi, Jong-Duck;Heu, Min-Soo;Kim, Jin-Soo
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.35 no.7
    • /
    • pp.926-934
    • /
    • 2006
  • The base for preparing oyster hydrolysate-added yogurt was consisted of whole milk (1,000 mL), skim milk (44.05 to 42.05 g), enzymatic oyster hydrolysates powder (OHP, 0 to 2.0 g) and pectin. The yogurt base was fermented with 7 kinds of starter cultures (3% based on yogurt volume), such as Lactobacillus acidophilus, lactobacillus bulgaricus, lactobacillus casei, Lactobacillus fermentum, Lactobacillus pentosus, Streptcoccus thermophilus and the mixed starters (L. bulgaricus and S. thermophilus) at optimal temperature. Processing condition and quality characteristics of the yogurt were evaluated by analyzing pH, titratable acidity, viscosity, viable cell count, functional properties and sensory evaluation. The results suggested that the optimal conditions for preparing the good quality yogurt revealed the mixed starters (L. bulgaricus and S. thermophilus) for starter culture, 1.0 g of 3 kDa hydrolysate for amount, and 5.5 hrs for fermentation time. The good quality yogurt showed 4.31 for pH, 1.07% for titratable acidity, 469 cps for viscosity and $4.9{\times}10^8\;CFU/mL$ for viable cell count. The hydrolysate-added yogurt was 2 times higher in ACE inhibitory and antioxidant activities than commercial yogurt, and kept good quality during storage of 15 days at $5^{\circ}C$.

The Effect of CO2 Fixation for Microalgae based on CO2 Concentration and Flow Rate (이산화탄소 농도 및 유속에 따른 하천 내 미세조류의 이산화탄소 고정 효과)

  • Park, Hyomin;Lee, Sangdon
    • Journal of Wetlands Research
    • /
    • v.20 no.4
    • /
    • pp.363-369
    • /
    • 2018
  • One of the recent environmental problems is climate change due to the increase of atmospheric $CO_2$, which causes ecological changes and various environmental problems. Therefore, various studies are being carried out to reduce $CO_2$ in the world in order to solve various environmental problems caused by increase of $CO_2$. The $CO_2$ reduction using microalgae is an environmentally friendly method by using photosynthesis reaction of microalgae. However, most studies using single species. There is no study on the $CO_2$ fixing efficiency of microalgae in natural rivers. Therefore, this study was to identify the microalgae in the Sum river and to analyze the growth characteristics of microalgae in the river to obtain optimal culture conditions. And the changes of biomass and chlorophyll-a of microalgae were analyzed according to $CO_2$ concentration and injection rate. The purpose of this study was to investigate the fixing efficiency of carbon dioxide in microalgae in natural rivers. Six kinds of dominant species were observed as a result of the identification of microalgae in Sum river(Ankistrodesmus falcatus, Scenedesmus intermedius, Selenodictyum sp., Xanthidium apiculatum var. laeve, Cosmarium pseudoquinarium, Dictyosphaerium pulchellum). All of these species were green algae. Biomass and chlorophyll-a increased with the increase of $CO_2$ concentration and biomass and chlorophyll-a increased faster flow rate at the same $CO_2$ concentration. Also, the quantity of $CO_2$ fixation on the microalgae tended to be higher when the flow rate of injected gas was faster. This study can be referred as being significant in the micro-algae in river. In addition, the optimal conditions for $CO_2$ fixation of microalgae in rivers and the quantification of the quantity of $CO_2$ fixation from microalgae in rivers can be used as basic data for future policy of $CO_2$ reduction.

Characteristic study and optimization of culture conditions for Bacillus amyloliquefaciens SRCM 100731 as probiotic resource for companion animal (Bacillus amyloliquefaciens SRCM 100731의 반려 동물용 프로바이오틱스 소재로서의 특성 규명 및 배양 조건 최적화)

  • Ryu, Myeong Seon;Yang, Hee-Jong;Jeong, Su-Ji;Seo, Ji Won;Ha, Gwangsu;Jeong, Seong-Yeop;Jeong, Do-Youn
    • Korean Journal of Microbiology
    • /
    • v.54 no.4
    • /
    • pp.384-397
    • /
    • 2018
  • The aim of this study is to screen the strains of Bacillus spp. possessing safety, probiotic activity, and so on, which can be utilized as probiotic resource for using the feed and supplement food of companion animal. About 300 isolates were isolated from traditional Korean sauces, four isolates that did not have or produce the six kinds of B. cereus type vomiting and diarrhea toxin genes, ${\beta}$-hemolytic, and three kinds of carcinogenic enzymes were selected. Antibiotic gene retention, cell surface hydrophobicity, antibiotic sensitivity, and glucose utilization were analyzed for four isolates, and finally SRCM 100731 was selected. SRCM 100731 was named as Bacillus amyloliquefaciens SRCM 100731 16S rRNA sequencing analysis, and carried out optimization of cell growth for industrial applications such as pet food and feed. The effects of 14 different components on cell growth were investigated and three significant positive factors, molasses, sodium chloride, and potassium chloride were selected as the main factors based on a Plackett-Burman design. In order to find out optimal concentration on each constituent, we carried out central composite design. The predicted optimized concentrations were 7% molasses, 1.1% sodium chloride, 0.5% potassium chloride. Finally, an overall about 7-fold increase in dry cell weight yield ($12.6625{\pm}0.0658g/L$) was achieved using the optimized medium compared with the non-optimized medium ($1.8273{\pm}0.0214g/L$). This research is expected to be highly utilized in the growing pet industry by establishing optimal cultivation conditions for industrial application as well as screening Bacillus amyloliquefaciens SRCM 100731 as probiotic resource for companion animal.

A interpretive Study of the Analects of Confucius's 'Ren(仁)' (『논어(論語)』의 '인(仁)'에 관한 해석학적(解釋學的) 연구(硏究))

  • Seo, Geun-sik
    • (The)Study of the Eastern Classic
    • /
    • no.36
    • /
    • pp.31-56
    • /
    • 2009
  • The core thought of Confucius("論語") is 'Ren(仁)'. Then, how ought we to interpret this 'Ren(仁)'? In this study, the researcher has interpreted 'Ren(仁)' from the perspective of Xiujizhiren(修己治人), which is the doctrine of Confucianism and its ideal. At first, the researcher closely reviewed Ren(仁) on the viewpoint of Xiuyang (修養). Ren(仁) is the most fundamental virtue that enables general populace to equip with their qualification as a human being. Specifically, to live like a human being, Ren(仁) is a must. That is to say, it will suffice if we only can expose well what was already cherished inside us, rather than exerting efforts to attain Ren(仁), in some contexts, that must achieve in order to live like a human being. The reason that we exert our efforts for self-cultivation is to bring this Ren(仁), which is foundation of human life, before the public. Even in relationship-building, Ren(仁) is necessary. Human being is not an existence that can live alone, but at all times, humans are required to build a relationship with others. To make this relationship-building lead into right direction, we need to think of that the standpoint of oneself and the other are identical. That is, when I myself and the other person are in the most optimal situation, then a right relationship-building can take place. This most optimal status is Ren(仁). The ideal of Confucianism is to establish a society where all people can enjoy their comfortable life. To accomplish such a society, each individual and society ought to be benevolent and to cherish humanity at the first place. That is to say, people should attain Ren(仁) from both aspects of Xiuji(修己) and Zhiren (治人). If Ren(仁) has not been attained from any of either side, then it is hard to say that the ideal of Confucianism is completely realized. However, Zhiren(治人) must be backed up by Xiuji(修己). For this reason, Kongzi(孔子) presented three steps in connection with this cultivation process, to wit, 'Cultivation of himself in reverential carefulness'(修己以敬) ${\rightarrow}$ 'Cultivation of himself so as to give rest to others'(修己以安人) ${\rightarrow}$ 'Cultivation of himself so as to give rest to all the people'(修己以安百姓). It is noticeable that Xiuji(修己) is included in all three phases. The society that Kongzi(孔子) longed for is still valid in this modern world. Therefore, Ren(仁) which was edified by Kongzi(孔子) is necessary for today's society. If we don't interpret Ren(仁) as with a fixed term lying stagnant in one place, then its definition shall be interpreted newly so as to suit the times and the situation of civil society, thus this Ren(仁) shall be the foundation for building a desirable society for humans.