• Title/Summary/Keyword: culture temperatures and media

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Isolation and in vitro culture of primary cell populations derived from ovarian tissues of the rockfish, Sebastes schlegeli

  • Ryu, Jun Hyung;Kim, Hak Jun;Bae, Seung Seob;Jung, Choon Goo;Gong, Seung Pyo
    • Fisheries and Aquatic Sciences
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    • v.19 no.2
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    • pp.9.1-9.7
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    • 2016
  • This study was conducted to identify the general conditions for the isolation and in vitro culture of ovary-derived cells in rockfish (Sebastes schlegeli). The effects of three different enzymes on cell retrieval from ovarian tissues were evaluated first, and then the ovary-dissociated cells were cultured under various culture conditions, with varying basal media and culture temperatures, addition of growth factors, and/or culture types. We found that collagenase type I treatment was effective for cell isolation from ovarian tissues. From a total of 42 trials to evaluate the effects of basal media and culture temperatures on cell culture of ovary-dissociated cells, we observed that Leibovitz's L15 medium was more supportive than Dulbecco's modified Eagle's medium for culture, and the cells could grow at all three temperatures tested, 15, 20, and $25^{\circ}C$, at least up to passage 2. However, growth factor addition did not improve cell growth. Introduction of suspension culture after monolayer culture expanded the culture period significantly more than did monolayer culture alone. Our results may provide a basis for developing an in vitro system for S. schlegeli germline cell culture, which will ultimately lead to improvement of the species.

Effects of Temperatures and Basal Media on Primary Culture of the Blastomeres Derived from the Embryos at Blastula Stage in Marine Medaka Oryzias Dancena

  • Choi, Jae Hoon;Gong, Seung Pyo
    • Journal of Embryo Transfer
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    • v.33 no.4
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    • pp.343-348
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    • 2018
  • Although the efforts to establish fish embryonic stem cells (ESCs) have been made for a long time, derivation of authentic ESCs that possess pluripotency is still difficult suggesting a need for the stepwise optimization of the methods to establish fish ESCs. Primary culture of the blastomeres from the embryos at blastula stage is a critical step for establishing continuous ESC lines. Here, we evaluated the effects of temperatures and basal media on primary culture of blastula embryo-derived blastomeres in marine medaka (Oryzias dancena). The blastomeres were isolated from the blastula embryos and cultured in various conditions designed by the combination of 4 temperatures including $28^{\circ}C$, $31^{\circ}C$, $34^{\circ}C$, and $37^{\circ}C$ and 2 basal media including Dulbecco's modified eagle's medium (DMEM) and Leibovitz's L-15 medium (L15). With the exception of a case cultured in L15 at $31^{\circ}C$, the rate of primary cell adherence reached 100% when the blastomeres were cultured over $31^{\circ}C$. The period for primary adherence was significantly shorter in the groups cultured in $34^{\circ}C$ and $37^{\circ}C$ than in the ones in $28^{\circ}C$ and $31^{\circ}C$. The proportion of subculture was significantly high in the group cultured in DMEM at $31^{\circ}C$ compared to the other groups. Collectively, we demonstrated that the culture in DMEM at $31^{\circ}C$ was effective to primary culture of the blastomeres derived from blastula embryos.

Studies on Culture of Leptospires On Growth of Leptospires in Cox's Liquid Media and Korthof's Media, Added Rabbit Serum and Thiamine Hydrochloride, Inactivated at High Temperature (Leptospira 균(菌) 배양(培養)에 관한 연구(硏究) -가열처리(加熱處理)한 토끼 혈청가배지(血淸加培地)와 Thiamine 첨가배지(添加培地)에서의 Leptospira 균(菌)의 발육(發育)에 관하여-)

  • Seo, Ik Soo
    • Korean Journal of Veterinary Research
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    • v.15 no.1
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    • pp.75-81
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    • 1975
  • The rate of growth of Leptospires were studied by culturing in Korthof's media and Cox's liquid media which were treated at four different temperatures. The effect of thiamine hydrochloride in the culture of leptospires were also studied. The results obtained were summerized as followings: 1. The best growth of Leptospira icterohaemorrhagiae and Leptospira canicola was observed in Korthof's media with the rabbit serum heated for 60 minutes at $56^{\circ}C$. 2. A good degree of growth of Leptospira icterohaemorrhagiae and Leptospira canicola was observed in both Cox's liquid media with rabbit serum heated for 30 minutes at $100^{\circ}C$ and with fresh rabbit serum. 3. The growth of Leptospira was enhanced by adding small amount of thiamine to the both media which were shown to be unsatisfactory for the growth of the organisms. 4. Culture of leptospires were attained by simply heating for 30 minutes at $100^{\circ}C$ both Korthof's media and Cox's liquid media containing rabbit serum obtained without aspetic procedures.

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Partial Characterization of an Anti-Listerial Bacteriocin from Enterococcus faecium CJNU 2524

  • Yang, Jung-Mo;Moon, Gi-Seong
    • Food Science of Animal Resources
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    • v.41 no.1
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    • pp.164-171
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    • 2021
  • Listeria monocytogenes is a representative foodborne pathogen and causes listeriosis. Enterococcus faecium CJNU 2524 was confirmed to produce a bacteriocin with anti-listerial activity. To establish optimal culture conditions for the production of the bacteriocin from E. faecium CJNU 2524, different media (MRS and BHI broth) and temperatures (25℃, 30℃, and 37℃) were investigated. The results showed that the optimal culture conditions were MRS broth and 25℃ or 30℃ temperatures. The crude bacteriocin was stable in a broad range of pH conditions (2.0-10.0), temperatures (60℃-100℃), and organic solvents (methanol, ethanol, acetone, acetonitrile, and chloroform). The bacteriocin activity was abolished when treated with protease but not α-amylase or lipase, indicating the proteinaceous nature of the bacteriocin. Finally, the bacteriocin showed a bactericidal mode of action against L. monocytogenes. Therefore, it can be a biopreservative candidate for controlling L. monocytogenes in dairy and meat products.

Comparison of Mycelial Growth Characteristics According to Culture Conditions of Ulleungdo Collection Strains (울릉도 수집 균주의 배양조건에 따른 균사 생장 특성 비교)

  • Kim, Minkyeong;Ahn, Chorong;Kim, Changmu
    • The Korean Journal of Mycology
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    • v.48 no.2
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    • pp.75-85
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    • 2020
  • The collection of biological data of indigenous species must comply with the Nagoya Protocol. Fungi contain various bioactive substances making them an attractive source of several products, including food and medicines. In this study, we investigated the growth characteristics of five indigenous fungal strains, Fomitiporia punctata, Polyporus ulleungus, P. brumalis, Gymnopus subnudus, and Tyromyces kmetii, isolated from samples collected in the Ulleungdo Island. The growth rates for each strain were assessed across various temperatures (20 ℃ to 35 ℃), culture media (Potato dextrose agar, Malt extract & Yeast extract agar, Malt extract agar, Malt extract & peptone agar, Sabouraud dextrose agar, and Modified Melin-Norkrans agar), and pH conditions (4.0 to 8.0). Additionally, we assessed the mycelial growth characteristics in liquid culture. The mycelial growth in different media varied across species; specifically, F. punctata (in MMNA), G subnudus (in MMNA), and P. brumalis (in MEPA) showed rapid growth. Optimal growth temperatures ranged between 25 ℃ and 30 ℃ for most species, with the exception of T. kmetii and P. brumalis, which were able to grow across all the temperatures tested. P. brumalis showed the best growth rate, whereas P. ulleungus showed the lowest growth potential. The optimal pH conditions for mycelial growth ranged between 4.0 and 5.0. In experiments using culture flasks, the dry weight of the culture filtrates decreased with the increasing incubation time and showed a significant decrease between 1 and 6 months of incubation, indicating that the five strains take longer than a month to fully use the culture media. Our findings highlight and establish the optimal growth conditions for five different fungal species that can be used in future application studies.

Mixed Culture Characteristics of Fungi Strains isolated from Korean Traditional Nuruk (전통누룩에서 분리한 양조용 곰팡이의 혼합 배양에 따른 품질 특성)

  • Mun, Ji-Young;Baek, Seong-Yeol;Ro, Hyeon-Su;Yeo, Soo-Hwan
    • Journal of the East Asian Society of Dietary Life
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    • v.27 no.3
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    • pp.348-363
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    • 2017
  • We analyzed the mixed culture characteristics of isolated brewing fungi obtained from Korean traditional nuruk for their development as fermentation starters. In a previous study, we selected appropriate fungi strains based on their large fungi bodies and strong enzyme activities. These fungi strains were mixed cultured under various combination of ratios, pHs and temperatures. As a result, total acidities of R. delemar mixed culture media were higher than those of A. oryzae cultured at $20^{\circ}C$ and $37^{\circ}C$. The pHs of A. luchuensis 34-1 and A. oryzae 37-7 as well as R. delemar 58-8 and A. oryzae 37-7 mixed culture media gradually increased with the culture period. On the contrary, total acidities of the mixed culture media gradually decreased. ${\alpha}-Amylase$ activities of R. delemar 26-4 and R. oryzae 82-7 mixed culture media were strong at $20^{\circ}C$ and pH 4, glucoamylase activities of R. delemar 58-8 and A. oryzae 37-7 were strong at $37^{\circ}C$, pH 3 and 1:2 ratio, and acidic protease activities of R. delemar 26-4 and A. oryzae 78-5 were strong at $20^{\circ}C$, pH 4 and 1:4 ratio. This study provides the optimal mixed culture conditions for fermentation starters based on changes in pH and total acidity, favorable enzyme activities, and fungi varieties.

Effects of Culture Temperatures and Media on Morphological Changes of Leptospira interrogans Isolated in Korea (한국에서 분리된 Leptospira의 배양조건에 따른 형태변화)

  • Lee, Tae-Yoon;Park, Jeon-Han;Uhm, Jeong-Ran;Lee, Bong-Ki;Lee, Won-Young;Kim, Joo-Deuk;Youn, Jung-Koo;Hahn, Kyu-Woung
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.3
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    • pp.209-218
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    • 1987
  • Leptospira interrogans, the causative organism of leptospirosis, is characterized by a fine helical morphology, and the helix is almost always right-handed. However, one of the striking features of recent isolates of L. interrogans in Korea was the heterogeneity in their morphology. Even under optimal culture conditions($30^{\circ}C$, EMJH medium), rods, spiral forms with right or left-handed helices, and even spherical forms of L. interrogans were present. Although the literature notes the presence of left-handed helices, long rods, and spherical forms in cultures of L. interrogans isolates, little is known about the cause of this morphologic heterogeneity. In an attempt to answer this question, this study was initiated to examine the effects of culture conditions, especially temperature and medium, on the morphology of L. interrogans. Four temperatures($5^{\circ}C$, $15^{\circ}C$, $30^{\circ}C$, and $37^{\circ}C$) and two types of media(Fletcher and EMJH) were used; one strain from Korean isolates and L. interrogans serovar canicola obtained from the Pasteur Institute(Paris, France) were employed throughout the study. The findings are as follows: 1. The L. interrogans isolated in Korea(UM-19) had a larger cell diameter($0.25{\sim}0.30\;{\mu}m$: $0.10{\sim}0.15\;{\mu}m$), and helix diameter($0.10{\sim}0.60\;{\mu}m$: $0.10{\sim}0.15\;{\mu}m$) than that obtained from the Pasteur Institute, but they varied in their distances between the helices($0.31{\sim}1.00\;{\mu}m$: $0.50{\sim}0.70\;{\mu}m$). 2, When UM-19 was grown at $37^{\circ}C$ after months or longer preincubation at $5^{\circ}C$ or $15^{\circ}C$, the majority of the organisms were spiral forms; however, they became rods when subcultured at $30^{\circ}C$ or $37^{\circ}C$. No significant morphological differences were found between Fletcher and EMJH media. 3. When L. interrogans serovar canicola was subcultured more than ten times at $37^{\circ}C$, some of the organism lost their motility as well as the hooks at either one or both ends, but only in Fletcher medium. The number of variants increased with the frequency of subculturing. These findings suggested that L. interrogans strain (UM-19) is different, in their morphology, from that of the Pasteur Institute, and its various morphologies may represent stages of the life cycle and vary with incubation temperature.

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Thermal Characteristics of Nutrient Solution and Root Media in Recycled Soilless Culture Systems (순환식 무토양재배시스템의 양액 및 배지의 온도변화 특성)

  • Son, Jung-Eek;Park, Jong-Seok
    • Journal of Bio-Environment Control
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    • v.7 no.1
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    • pp.71-77
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    • 1998
  • The root-zone environment is an important factor to the plant growth and it is closely related to the thermal characteristics of the root media. In this study thermal characteristics of root media with ambient environmental conditions were analyzed. The temperatures of nutrient solution as well as inside air of culture bed were measured in Nutrient Film Technique(NFT) and Deep Flow Technique(DFT) systems, and also the temperatures of root media measured in aggregate culture systems , The temperature of nutrient solution of NFT system with as low as 3$\ell$/min of flow rate was 3$^{\circ}C$ higher than that with 5 $\ell$/min of flow rate in the daytime, and the temperature of inside air was 2$^{\circ}C$ higher at night. And the temperature of nutrient solution of DFT system with as low as 0.8 cm of water level was 1-2$^{\circ}C$ higher than that with 1 8 cm in the daytime, and the temperature of inside air was almost same at night. The root-zone temperatures in the perlite and rockwool granulate systems with film mulching were 3$^{\circ}C$ higher than those without film mulching in the daytime. However, the rockwool slab system with film mulching showed the same trend as rockwool granulate system, but relatively higher temperature than any other medium because of the exposure of media surface to the ambient air. Additionally the temperature below the plant was measured 3$^{\circ}C$ lower than that between plants.

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Growth characteristics and lipid content of three Korean isolates of Botryococcus braunii (Trebouxiophyceae)

  • Lee, Chan-Hee;Chae, Hyun-Sik;Lee, Seung-Hoon;Kim, Han Soon
    • Journal of Ecology and Environment
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    • v.38 no.1
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    • pp.67-74
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    • 2015
  • Three strains of the green microalga Botryococcus braunii (JJS, KCM, and KJD) were isolated from different water bodies in Korea and grown as batch cultures in the laboratory. The effects of different growth media and temperatures on the growth rate were investigated, as well as the effect of temperature on the total lipid content and lipid profile. All three strains had the highest growth rates in BG-11 medium and at $25^{\circ}C$. Maximal lipid production ($gL^{-1}$) was at $30^{\circ}C$ in the JJS strain and at $25^{\circ}C$ in the KCM and KJD strains. However, all the three strains produced the greatest percent dry weight of total lipids at $15^{\circ}C$ and had the lowest percent dry weight of total lipids at $25^{\circ}C$. In general, oleic acid, linolenic acid, and behenic acid were the most common fatty acids in all three strains. However, the three strains varied considerably in their fatty acid profiles at different culture temperatures.

Influences of DNP and Low Temperature on the Promotive Action of Calcium in Oenothera Semi-Vitro Pollen Growth (In Semi-Vitro 달맞이꽃분화의 석탄에 의한 생장촉진과 DNP 및 저온의 그 억제작용에 관하여)

  • 정병화
    • Journal of Plant Biology
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    • v.12 no.2
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    • pp.23-27
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    • 1969
  • The promotive effect of Ca in semi-vitro culture of Oenothera biennis pollen was by and large identical to that of in vitro systems. This promotive effect became additive when boron was supplemented to the media and even more so if K was further supplemented. No such Ca action was observed then performed under the conditions of relatively low temperature. An uncoupling agent of ATP in respiration, DNP, inhibited the promotive action of B, but not that of Ca. The inhibitory effect of DNP was greater at low temperatures. IAA was rather inhibitory on pollen growth in semi-vitro culture, even much greater than it if DNP was supplemented to the IAA-containing media.

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