• Parasites, Hosts and Diseases
• /
• v.37 no.2
• /
• pp.121-125
• /
• 1999

Inoculation of human fecal cysts to suckling Mongolian gerbils, two Giardia lamblia isolates, Kl and K2, were established as axenic cultures. Using this in vitro culture, both two Giardia isolates were grouped by using two genetic analysis. With genetic analysis of SSS-rDNA sequences, both K1 and K2 were found as members of hopkins'group 1, despite some nucleotide differences noticed in K1 (5 differences/292 bases.). The other genetic study used PCI-RFLP of the tim (triose phosphate isomerase) Nash's group 2 can bot be a separate group, but a part of Hopkins' group 1.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.8 no.3
• /
• pp.165-168
• /
• 1988

This study was conducted at the rice field of Livestock Experiment station, Suweon to investigate the effects of harvesting time on forage production and quality of rye (Secale cereale L.) on paddies. The results obtained are summarized as follows; 1. Fresh matter yield was peaked at the 10 days (May 10) after heading, however, dry matter yield was increased in proportion to maturity. 2. Digestibility (in vitro) showed the 83-77% by heading stage (Apr. 25-30) but decreased to 66-58% after heading stage. 3. Digestible yield reached to maximum level, 619.2 kg/lOa at the 10 days after heading (May 10). 4. The content of crude protein and crude fat in forage rye was decreased with advancing the maturity. 5. The lactic acid content of rye silage ranged from 1.3% to 2.0%.

• Proceedings of the Korean Society of Organic Agriculture Conference
• /
• 2009.12a
• /
• pp.316-316
• /
• 2009

This study was conducted to investigate the effects of environment-friendly materials on garlic cultivation, and the methods for processing garlic at proper ripening stage to increase its antioxidant properties and market value. The functional properties of garlic and processed black garlic were analyzed in vitro and ex vivo. The ethanol extracts of garlic and black garlic were used in the antioxidant, linoleic acid autooxidation, lipid oxidation and RAW264.7 indicator cell using NO induced activity analyses. Also, toxicity tests by RBL-2H3 and RAW264.7 indicator cell using MTT assay were compared. Obesity inhibition test of garlic and black garlic were also studied.

• Korean Journal of Medicinal Crop Science
• /
• v.12 no.6
• /
• pp.448-453
• /
• 2004

Transformed roots of Schisandra chinensis were obtained following co-cultivation of in vitro cultivated plantlet segments with Agrobaterium rhizogens ATCC15834. This root was examined for its growth and gomisin J contents under various culture conditions. Among the six basal culture media tested, WPM (Lloyd & McCown, 1980) medium supplemented with 5% sucrose was the best roots growth 6.2 (g D.W/flask) and gomisin J accumulation 1.56 $(X10^{-3}\;ug/g\;D.W)$. Initial inoculum size correlated with the yield of biomass while gomisin J contents was not affect. Gomisin J production was influenced by the initial sucrose concentration and the highest production yield was achieved at the concentration of 7%. The optimal shaking speeds for roots growth and gomisin J production was 120 and 140 rpm, respectively.

• Journal of Plant Biotechnology
• /
• v.35 no.1
• /
• pp.63-68
• /
• 2008

Single-node stem pieces ca. 1 cm in length containing a axillary bud were obtained from in vitro plants of potato (Solanum tuberosum L.). The influences by a position of the node and the existence of a leaf at the node were observed in the single-node culture on the 8% sucrose MS medium. The effect of CCC was also investigated for the microtuberization. The apical part node was excellent in the tuberization not to mention shoot length, fresh weight, diameter, the number of node on the in vitro culture of a single-node than the lower part. The differences in the diameter of a tuber formed in the part of the axillary bud on all treatments including the cultivation of the apical part node were not recognized. However, the fresh weight of the tuber showed high value in the tuber formed at the axillary bud of shoot apex part. At 20 days after cultivation, tuberization was promoted in the new stolen that developed from the bud of node with a leaf under SD condition of 8 hours at $20^{\circ}C$. The tuberization from axillary bud of the single-node without leaf was inhibited at high temperature of $28^{\circ}C$ regardless of daylength. Whereas, tuberization at $20^{\circ}C$ and $28^{\circ}C$ was similar without the difference under SD condition but the tuber formation ratio were low. CCC 500 mg/L promoted tuberization and the effect was also showed even under LD condition at $28^{\circ}C$. The inhibiton of tuberization under LD and high temperature condition could be solved by treatment with CCC.

• Parasites, Hosts and Diseases
• /
• v.27 no.4
• /
• pp.231-248
• /
• 1989

The growth and development of the metacercariae of F. seoulensis cultivated in vitro or on the chick chorioallantois were assessed by comparison with the optimum process of maturation in albino rats and new born chickens. The process of maturation was divided for convenience into six stages: Stage 1 ; cell multiplication, Stage 2; body shaping, Stage 3; separation of genital anlagen, Stage :1 organogeny, Stage 5; gametogony, and Stage 6: oviposition. In Hank's and Tyrode's .solutions, the metacercariae were alive up to 200 days or more at $4^{\circ}C$ without any development. The in vivo maturation process in rats or chicks was as follows: stage 1 from 6 hours; stage 2 from 24 hours; stage 3 from 48 to 72 hours; stage 4 from 3 to 4 days; stage 5 from 4 to 5 days; and stage 6 from 5 to 8 days. Despite unsuccessful infection of the metacercariae to 12 day old chicks, fully mature worms of stage 5 or 6 were recovered from new born chicks (1 to 2 days old), The metacercariae of F. seoulensis grown in vitro were up to stage 3 and no further maturation was observed. Of various media employed, the medium NCTC 109 (Gibco) or NCTC 135(Gibco) supplemented with 20% egg yolk or 20% whole egg macerate or 0.5% yeast was basically required for the earlier development of the fluke. It took 16.1 days(in average) to reach the stage 3 after cultivation. The metacercariae cultivated on the chorioallantoic membranes of 6∼13 day old chick embryo at 37∼38℃ showed their full development up to stage 5 or 6. However, the worms were in general remarkably retarded, compared with those grown in rats or chickens. In the experiments of worm transplant, although the transfer was failed from in vitro culture to in vivo of rats(Per os), the transplants from in vitro culture to the chorioallantois and from the choriollantois to in vivo of rat host were successful with or without development of the transferred worms. In the present study, it was observed that the metacercariae of F, seoulensis can be maintained in vitro media with poor development as well as fully matured in 1 to 2 day-old chicks or on the chorioallantois at a very low rate.

• Korean Journal of Korean Medical Institute of Dermatology and Aesthetics
• /
• v.1 no.1
• /
• pp.53-90
• /
• 2005

Purpose: Contents of astragaloside I, II and IV, cytotoxicity, anticancer activity, immunomodulatory activity and antioxidant capacity were to be compared as a function of the cultivated years as one, three, five and seven years. Method: Major components of Astragali membranacei Radix were separated as astragaloside I, astragaloside II, astragaloside IV by HPLC analysis. Cytotoxicity and anticancer activities were measured by MTT and SRB assay. For immunomodulatory activity, the secretion of IL -6 and $TNF-{\alpha}$, NK cell activation and macrophage activation were observed as well as kinetics of responding to human T cells by a microphysiometer. In vitro antioxidant activities were measured by several radical scavenging activities of superoxide anion radican, DPPH, LDL and linoleic acid. For in vivo activity, the activation of SOD, GSH-px, catalase, ALDH and ADH was measured as well the relative weight of liver. Result : 1. For HPLC analysis, the contents of all of astragaloside I, astragaloside II, astragaloside IV were in order of three, five, one and seven years. 2. The cytotoxicity of normal human lung cell line, HEL299 showed lower than 18% in adding 0.25 mg/ml, and 28.9% in adding 1.0 mg/ml of water extract of seven year root. For methanol extracts, three year root showed highest cytotoxicity as 35.2 % and there was no difference between the cultivated years. 3. For anticancer activities, methanol extracts of one and three year roots showed relatively high inhibition of human stomach cancer cells, AGS, breast cancer cells, MCF-7, lung cancer cells, A549 and liver cancer cell, Hep3B as well as high selectivities. 4. The water extract of seven year root could yield high secretion of IL-6 from both human Band T cells while the methanol extracts of three and five year roots secreted high amounts of IL-6 and $TNF-{\alpha}$ from both Band T cells. 5. As a result of in vitro antioxidant activities, both water and methanol extracts from five and seven year roots showed high activities for superoxide anion radical scavenging activity, inhibiting linoleic acid peroxide and contents of total phenols. 6. For in vivo tests, Mn-SOD and GSH-px activities and weight of liver were better in adding seven year root. For ALDH activity one year root was better and for ADH activity five year root. Overall speaking, seven year root showed relatively better antioxidant activities. Conclusion:There was difference of the contents of astragaloside I, astragaloside II, astragaloside IV according to cultivation year. Methanol extract showed better activities of anticancer and immune activation rather than water extract Interestingly enough, for methanol extracts, overall activities were improved as the cultivation year increased. There might be further investigation required for the clinical uses of the results as several biological activities varied according to the cultivated year of Astragali membranacei Radix.

• Journal of Animal Science and Technology
• /
• v.64 no.6
• /
• pp.1105-1116
• /
• 2022

Recently, we reported the robust in vitro three-dimensional (3D) expansion of intestinal organoids derived from adult bovine (> 24 months) samples. The present study aimed to establish an in vitro 3D system for the cultivation of intestinal organoids derived from growing cattle (12 months old) for practical use as a potential alternative to in vivo systems for various purposes. However, very few studies on the functional characterization and 3D expansion of adult stem cells from livestock species compared to those from other species are available. In this study, intestinal crypts, including intestinal stem cells, from the small intestines (ileum and jejunum) of growing cattle were isolated and long-term 3D cultures were successfully established using a scaffold-based method. Furthermore, we generated an apical-out intestinal organoid derived from growing cattle. Interestingly, intestinal organoids derived from the ileum, but not the jejunum, could be expanded without losing the ability to recapitulate crypts, and these organoids specifically expressed several specific markers of intestinal stem cells and the intestinal epithelium. Furthermore, these organoids exhibited key functionality with regard to high permeability for compounds up to 4 kDa in size (e.g., fluorescein isothiocyanate [FITC]-dextran), indicating that apical-out intestinal organoids are better than other models. Collectively, these results indicate the establishment of growing cattle-derived intestinal organoids and subsequent generation of apical-out intestinal organoids. These organoids may be valuable tools and potential alternatives to in vivo systems for examining host-pathogen interactions involving epithelial cells, such as enteric virus infection and nutrient absorption, and may be used for various purposes.

• 한국생물공학회:학술대회논문집
• /
• 2003.04a
• /
• pp.302-305
• /
• 2003

It is difficult to obtain sufficient healthy skin for coverage of a wide area of skin wound. In the skin, an additional population of living epithelial cells is located in the outer root sheath (ORS) of hair $follicles.^{1),2)}$ ORS cells should be a good source of epithelium because they are easily obtainable and patients do not have to suffer from scar formation at donor sites. We modified ordinary primary culture technique for the purpose of solving such problem that epithelial cells have a low propagation and easy aging during culture periods. First of all, we improved primary cultivation methods. In the ordinary primary culture, average yield of human ORS cells was $2\;{\times}\;10^3$ cells/follicle by direct incubation with trypsin (0.1%)/EDTA (0.02%) solution for 15 min at $37^{\circ}C$ but we could obtain about $6.5\;{\times}\;10^3$ cells/follicle by two step enzyme digestion method with dispase (1.2 U/ml) and trypsin (0.1%)/EDTA (0.02%) solution. So we could achieve three times higher primary cultured ORS cell yield. Secondly, we could obtain total $2\;{\times}\;10^7$ cells in serum free medium and even more total $6\;{\times}\;10^7$ cells in modified E-medium with mitomycin C-treated feeder cells during 17 days. Using the cultured ORS cells, and we could make bioartificial skin equivalent in vitro and concluded that ORS cells were progenitor cells for skin epithelial cell.

• Journal of Microbiology and Biotechnology
• /
• v.30 no.11
• /
• pp.1785-1791
• /
• 2020

In a previous study, the sequential optimization and regulation of environmental parameters using the PhotoBiobox were demonstrated with high-throughput screening tests. In this study, we estimated changes in the biovolume-based composition of a polyculture built in vitro and composed of three algal strains: Chlorella sp., Scenedesmus sp., and Parachlorella sp. We performed this work using the PhotoBiobox under different temperatures (10-36℃) and light intensities (50-700 μmol m^-2 s^-1) in air and in 5% CO₂. In 5% CO₂, Chlorella sp. exhibited better adaptation to high temperatures than in air conditions. Pearson's correlation analysis showed that the composition of Parachlorella sp. was highly related to temperature whereas Chlorella sp. and Scenedesmus sp. showed negative correlations in both air and 5% CO₂. Furthermore, light intensity slightly affected the composition of Scenedesmus sp., whereas no significant effect was observed in other species. Based on these results, it is speculated that temperature is an important factor in influencing changes in algal polyculture community structure (PCS). These results further confirm that the PhotoBiobox is a convenient and available tool for performance of lab-scale experiments on PCS changes. The application of the PhotoBiobox in PCS studies will provide new insight into polyculture-based ecology.