• Title/Summary/Keyword: agar-agar

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Development of a Three-dimensional Hydrogel System for the Maintenance of Porcine Spermatogonial Stem Cell Self-renewal

  • Park, Ji Eun;Park, Min Hee;Kim, Min Seong;Yun, Jung Im;Choi, Jung Hoon;Lee, Eunsong;Lee, Seung Tae
    • Journal of Embryo Transfer
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    • v.32 no.4
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    • pp.343-351
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    • 2017
  • Porcine spermatogonial stem cells (SSCs) prefer three-dimensional (3D) culture systems to 2D ones for the maintenance of self-renewal. Of the many 3D culture systems, agar-based hydrogels are candidates for supporting porcine SSC self-renewal, and there are various types of agar powder that can be used. In this study, we sought to identify an agar-based 3D hydrogel system that exhibited strong efficacy in the maintenance of porcine SSC self-renewal. First, 3D hydrogels with different mechanics were prepared with various concentrations of Bacto agar, lysogeny broth (LB) agar, and agarose powder, and the 3D hydrogel with the strongest alkaline phosphatase (AP) activity and greatest increase in colony size was identified for the different types of agar powder. Second, among the porcine SSCs cultured in the different 3D hydrogels, we analyzed the colony formation, morphology, and size; AP activity; and transcription and translation of porcine SSC-related genes, and these were compared to determine the optimal 3D hydrogel system for the maintenance of porcine SSC self-renewal. We found that 0.6% (w/v) Bacto agar-, 1% (w/v) LB agar-, and 0.2% (w/v) agarose-based 3D hydrogels showed the strongest maintenance of AP activity and the most pronounced increase in colony size in the culture of porcine SSCs. Moreover, among these hydrogels, the strongest transcription and translation of porcine SSC-related genes and largest colony size were detected in porcine SSCs cultured in the 0.2% (w/v) agarose-based 3D hydrogel, whereas there were no significant differences in colony formation and morphology. These results demonstrate that the 0.2% (w/v) agarose-based 3D hydrogel can be effectively used for the maintenance of porcine SSC self-renewal.

A Manufacturing Technique of Agar with Strong Gelling Ability from Gelidium amansii (우뭇가사리로부터 고강도 한천의 제조)

  • DO Jeong-Ryong;PARK Jin-Hee;JO Kil-Suk
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.5
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    • pp.673-676
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    • 1998
  • Preparative conditions of high-gel strength agar from Gelidium amansii have been studied, The effect of NaOH pretreatment on the quality and yields of agar extracted from Gelidium amansli was examined. The Bel strength of agar extracted from C. amansii pretreated with NaOH was higher than that of agar extracted from G. amansii non-pretreated with NaOH. The gel strength of agar extracted from G. amansii was influenced by concentration, temperature and time of pretreatment with NaOH. It was found that the proper concentration, temperature and time of NaOH pretreatment to produce high-gel strength agar was $6\%$ NaOH, $80^{\circ}C$ and 2$\~$3 hrs. The principal sugars of agar extracted from G. amansli were galactose and 3,6-anhydrogalactose.

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Extraction and Purification of Agar from Gelidium amansii (우뭇가사리로부터 한천의 추출 및 정제)

  • Do Jeong-Ryong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.3
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    • pp.423-427
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    • 1997
  • The effect of different treatments on the quality and yield of purified agar produced from Gelidium amansii has been studied, and the extraction condition ol agar produced from G. amansii has been examined. The contents of ash, sulfate in agar produced from g. amansii collected from different places were $2.63\~2.92\%\;and\;1.38\~1.78\%$, respectively. Yields and gel strength of agar produced from G. amansii collected from different places were $31.6\~46.8\%\;and\;496\~887g/cm^2$, respectively. It was effective to extract agar at $120^{\circ}C\;for\;2\~3hrs$. Agar was purified by D.W. washing, EDTA washing, chitosan treatment, CPC treatment, PEG treatment, ethanol precipitation, acetone precipitation and propanol precipitation. The mineral contents of agar produced from Gelidium amansii were Na (2934ppm), Ca (2472ppm), Mg (2259ppm), K (2527ppm), P1(81.1ppm), Fe (66.4ppm), Al (71.7ppm), Zn (29.7ppm) and Pb (ND: not detected), respectively.

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Comparison of Four Different Isolation Media for Staphylococcus aureus (황색포도상구균 분리배지 비교)

  • Oh, Min-Hee;Kang, Seong-Il;Hong, Sang-Phil;Oh, Se-Wook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.5
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    • pp.606-611
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    • 2009
  • Performance test was carried out between selective media which are generally used in Staphylococcus aureus isolation from food. Sensitivity, determined according to the appearance of characteristic colonies when 30 different S. aureus strains were tested, resulted as Baird-Parker agar (RPF)> $Petrifilm^{TM}$ Staph Express plate> Baird-Parker agar> Mannitol salt agar. Also, the four different media showed the same selectivity because all tested media did not produce the false positive colonies. Recovery efficiency from the artificially inoculated foodstuff was almost the same for the tested media. Presumptive colonies were collected from the dried fishery product using Mannitol salt agar and collected strains were tested on 4 different selective agar. Almost presumptive strains did not show the false positive colonies except for S. carnosus ssp carnosus. This strain was identified as false positive colonies on Mannitol salt agar, Baird-Parker agar and $Petrifilm^{TM}$ Staph Express plate. But Baird-Parker agar (RPF) did not show the false positive colonies with the same strains. So, it was concluded that the Baird-Parker agar (RPF) has more higher selectivity than other tested media in this experiment.

Morphological and Cultural Characters of Didymella bryoniae on Seeds and Culture Media (종자(種子) 및 배지상(培地上)에서의 오이류(類) 덩굴마름병균의 형태적(形態的) 및 배양적(培養的) 특징(特徵))

  • Lee, Du-Hyung
    • The Korean Journal of Mycology
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    • v.10 no.1
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    • pp.7-13
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    • 1982
  • Habit characteristics of imperfect and perfect stage of D. bryoniae encountered on naturally infected seeds of cucumber and pumpkin were studied by the blotter method and compared with those grown on Difco potato dextrose agar (PDA), V-8 juice agar and water agar leaf medium (WALM). Most of the pycnidiospores obtained from each isolate of this fungus grown on PDA were non-septate and microtype. Non-septate pycnidiospores were predominanted in all isolates, but a macrotype of the non-septate and a number of uniseptate pycnidiospores were produced on V-8 juice agar and water agar leaf medium. On seed the pycnidiospores were mostly non -septate, but rarely uniseptate ones were also found. On radicle of cucumber seed, the pycnidiospores were non-septate and uniseptate but small percentage biseptate with somewhat constricted at septa. Pycnidiospores produced on V-8 juice agar and water agar leaf medium were similar to those produced on seeds. In the present investigation the perithecia were mostly globose to subglobose with apical papillate ostiole and whitish spore masses formed on the ostiole of perithecia, either on naturally infected seed or on culture media. The mature perithecia were dark brown to black. They were partially embedded or erumpent on seed coat and culture media. The perithecia varied in size within a much narrower range than the pycnidia. But perithecial formation of this fungus on PDA, V-8 juice agar, WALM and seed varied considerably depending upon isolate and culture media.

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Formation of Furans during the Acid Hydrolysis of Agar and Their Removal by Treatments of Lime, Steam-stripping and Hydrophobic Resins (한천의 산 당화에 의한 Furan화합물의 생성 및 제거)

  • Kim, Na-Hyun;Lee, Jae-Won;Seo, Yung-Bum;Yoon, Min-Ho
    • Korean Journal of Agricultural Science
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    • v.36 no.2
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    • pp.225-232
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    • 2009
  • The ratio of saccharification and formation of furans during the acid hydrolysis of agar with oxalic acid and sulfuric acid were examined base on the contents of the agar and acids. The ratio of saccharification in oxalic acid appeared to be 51~59% somewhat higher than 49~61% of sulfuric acid. Formation of the furans during the acid hydrolysis increased proportional to the contents of agar and acid. The relative formation ratio was high 10~47% for furfural (FUR) and 15~29% for hydroxy-methyl furfural (HMF) in 0.5~1.25% sulfuric acid rather than those of oxalic acid. When comparing the removal efficiency of the furans using an alkali treatment, steam stripping and hydrophobic resins, FUR was eliminated 60% by the alkali treatment, 62~90% by steam stripping and 71~75% by Amberlite XAD4 and 7HP, while HMF was removed to low levels of 10.5%, 4~17% and 13~25%, respectively. The loss of reducing sugar was also observed in process of the removal of furans, and the loss rate was the level of 2~4% in alkali treatment, 11~16% in steam stripping and 7~9% in Amberlite resins.

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Development of Modified Selective Media to Differentiate Cryptococcus Species Complex and its Serotypes using Natural Materials

  • Park, Gyu-Nam;Kim, Hye-Ran;An, Dong-Jun;Chae, Hee-Sun;Chang, Kyung-Soo
    • Biomedical Science Letters
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    • v.23 no.2
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    • pp.64-72
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    • 2017
  • The formation of brown colonies due to phenol oxidase activity on classic agar media containing natural material extracts of Helianthus annuus or on medium containing L-3,4-dihydroxyphenylalanine has been used to identify Cryptococcus species complex. In this study, various natural materials were used to develop a modified medium and to identify five major serotypes of Cryptococcus species complex. Serotypes A, D, and A/D were pigmented on medium using Perilla frutescens var. japonica Hara (PerJ agar) after a three-day incubation. Serotypes B and C were pigmented on PerJ agar after four- and five-day incubations, respectively. Growth time and pigmentation of the five serotypes occurred more rapidly on PerJ agar than on the other media. In addition, colony morphology, size, and pigmentation were specific by serotype. In conclusion, PerJ agar should be used in clinic settings to identify Cryptococcus species complex and its serotypes rapidly.

Occurrence of severe soybean-sprout rot caused by Pythium deliense in the recirculated production system

  • Yun, Sung-Chul
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.92.2-93
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    • 2003
  • Severe soybean-sprout rot was found at the mass productive factory in 2000 and 2001 and it caused 10-20% loss of the production. Pythium sp. was isolated almost 90% by potato dextrose agar from rotted root and hypocotylsof the sprouts. And the pathogencity tests using test tubes with 2% water agar and small containers (30 ${\times}$ 30 ${\times}$ 50 cm, WxLxH) cultivation were shown a similar rot on roots and hypocotyls. The fungal mycelium grew rapidly on the water agar and it prevented the seed germination. Density of the Pythium sp. in the recycled water system at the factory was periodically measured using a selective medium, corn meal agar with Pimaricin 10 mg, Rifampicin 10 mg, Ampicillin 100 mg per 1 liter in order to check the contamination of recycled water. After fitering step using 5 and 1 ml in the recycled system was applied and it was effectively controlled Pythium rot. The daily yield of sprout was stable and the occurrenceof Pythium in the recycled water was much less after filtering. The fungal isolates were identified as Pythium deliense Meurs based on various mycological characteristics on corn meal agar and sucrose-asparagine bentgrass leaf culture medium. P. deliens oogonia were spherical, smooth, 19-23 urn in diameter, and their stalk bending toward antheridia. Antheridia were straw hat-shaped, curred club-shaped, therminal or intercalary, monoclinous, occasionally diclinous, 12∼15 ${\times}$ 8∼11 um, 1(∼2) per oogonium.

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Use of Sucrose-Agar Globule with Root Exudates for Mass Production of Vesicular Arbuscular Mycorrhizal Fungi

  • Thangaswamy Selvaraj;Kim, Hoon
    • Journal of Microbiology
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    • v.42 no.1
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    • pp.60-63
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    • 2004
  • A sucrose-agar globule (SAG) was newly introduced to increase production of the vesicular arbuscular mycorrhizal (VAM) fungal spores, Gigaspora gigantea and Glomus fasciculatum. An SAG inoculum and a sucrose-agar globule with root exudates (SAGE) inoculum were prepared, and their spore productions were compared with a soil inoculum. When the SAGE was used as the inoculum on sucrose-agar medium plates the number of spores was increased (35% more than the soil inoculum). After the soil inoculum and SAGE were inoculated on an experimental plant, Zingiber officinale, the percentage root colonization, number of VAM spores, and dry matter content were analyzed. It was observed that the SAGE showed a higher percentage of root colonization (about 10% more), and increases in the number of spores (about 26%) and dry matter (more than 13%) for the two VAM fungal spores than the soil inoculum. The results of this study suggested that the SAGE inoculum may be useful for the mass production of VAM fungi and also for the large scale production of VAM fungal fertilizer.

Diversity of Cultured and Uncultured Bacteria in the Gut of Olive Flounder Paralichthys olivaceus (넙치(Paralichthys olivaceus) 장관의 배양 및 비배양 방법에 의한 세균의 다양성)

  • Kim, Ahran;Kim, Do-Hyung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.48 no.4
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    • pp.447-453
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    • 2015
  • We determined the optimal culture conditions for obtaining the maximum number of intestinal bacteria from the olive flounder Paralichthys olivaceus, and studied bacterial diversity using both culture-dependent and culture-independent methods. Using six culture conditions, mean bacterial numbers were greater than $10^6$ per gram of gut mucus, regardless of the medium. However, the bacterial diversity, based on colony morphology, appeared much higher on Marine agar (MA) and Zobell 2216 agar than on other media. We found eight and 17 cultured bacterial phylotypes with 99% minimum similarity in gut mucus grown on MA and tryptic soy agar, respectively. Furthermore, we used genomic DNA extracted from gut mucus to generate 78 random clones, which were grouped into 25 phylotypes. Of these, six were affiliated with Firmicutes, Actinobacteria, and Verrucomicrobia, and were not found using our culture-dependent methods. Consequently, we believe that Marine agar and Zobell 2216 agar are optimal media for culturing diverse intestinal microbes; we also discovered several novel sequences not previously recognized as part of the gut microbiota of olive flounder.