• Parasites, Hosts and Diseases
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• 제37권2호
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• pp.121-125
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• 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.

• 한국초지조사료학회지
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• 제8권3호
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• pp.165-168
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• 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%.

• 한국유기농업학회:학술대회논문집
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• 한국유기농학회 2009년도 하반기 학술대회
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• pp.316-316
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• 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.

• 한국약용작물학회지
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• 제12권6호
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• pp.448-453
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• 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
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• 제35권1호
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• pp.63-68
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• 2008

감자의 마디배양에서 이식하는 마디의 위치에 따라 기내 소괴경 형성에 미치는 차이와 배양하는 마디조직체에서의 잎의 유무가 기내소괴경의 형성에 미치는 온도, 일장 및 CCC의 영향을 검토하였다. 마디의 위치에 따른 감자의 생육에 있어서는 초장, 생체 중, 직경, 마디수는 물론 괴경형성에 있어서도 정단부위가 하부부위보다 양호하였다. 액아의 부위에서 형성된 소괴경의 직경에 있어서는 정단부를 포함한 모든 처리구에서 큰 차이를 확인할 수 없었으나 괴경의 생체중은 정단부위의 액아에서 형성된 괴경에서 높은 수치를 보였고 다른 마디에서는 개체간의 큰 차이가 없었다. 잎의 부착 유무와 온도와 일장에 따른 괴경형성에 있어서는 잎이 부착된 마디를 $20^{\circ}C$의 단일조건에서 배양할 때 양호하였으나, 고온에서는 괴경형성이 일장에 관계없이 저조하였다. 잎이 부착되지 않은 마디에서의 괴경형성은 저온과 고온 모두 단일조건에서 양호하였다. CCC처리에 의한 효과에 따른 감자의 생육에 있어서 초장, 생체중, 직경, 마디수는 CCC를 첨가한 배지에서 양호하였고 특히 일장 처리가 길어질수록 양호한 결과를 보였다. 괴경형성을 촉진시키는데 일장이 짧을수록 양호하였고, CCC를 첨가한 배지에서 양호한 효과를 보였다.

• Parasites, Hosts and Diseases
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• 제27권4호
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• pp.231-248
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• 1989

Fibricola seoulensis의 피낭유충을 in vitro 및 닭 장뇨막(chorioallantoic membrane)에서 배양하고 그 발육 및 성장을 흰쥐 및 갓난 병아리에서의 정상 성숙 과정과 비교하였다. 성숙 과정을 충체 염색 전체 표본과 절편 표본에서 현미경적으로 관찰하고 편의상 다음 여섯 시기로 구분하였다. 제 1시기 : 세포 증식, 제2시기 : 체형의 구성, 제 3시기 . 생식원기의 분리, 제 4시기 : 기된 형성기, 제 5시기 : 생식 분열기, 제 6시기 : 산란기 1. Hank's 및 Tyrode액에 $4^{\circ}C$에 보존한 피낭유충은 아무런 발육없이 200여일 이상 생존할 수 있음이 확인되었다. 2. 흰쥐 및 병아리 숙주 내(in vive)에서의 성숙 과정을 실험 감염 후 6시간에서 9일 사이에 회수한 충체의 발육 시기를 결정함으로써 경시적으로 관찰한 바 정상 숙주인 흰쥐에서 감염 후 6시간에서 tribocytic organ 내 선 세포의 수 증가가 인정되 었고(제 1시기), 24시 간에서 생식 활기의 증대와 전후 양부의 충체 형성이 시작되 었으 며(제 2시기), 48~72시간에서 생식 원기 세포 군의 분리가 인식되었다(제 3시기). 감염 후 3~4일째에서는 뚜 렷한 전후 층체의 구분과 난소 및 고환의 은과이 형성되기 시작하였으며(제 4시기), 4~5일째 충체에서는 자웅 생식기 및 난형성이 출현되었고(제 5시기), 5~8일째 충체에서는 산란을 인정할 수 있었다(제 6시기). 부화 12일째 된 병아리 실험 감염에서는 충체를 회수할 수 없었으나 112일 된 갓난 병아리에서는 제 5 내지 제 6시기로 인정되는 성숙된 충체를 회수하였다. 갓난 병아리에사의 피낭유충 실험 감염 후 성숙 과정은 정상 숙 주 내에서와 거의 유사하였다. 3. In vitro 내에서의 F. seoulensis 피낭유충은 완전한 성숙 과정을 관찰할 수 없었으나 생식 원기 세포 군의 분리를 관찰할 수 있는 제 3시기 까지의 발육 및 성장이 인정되었다. In vitro 배양 총체를 배양 전 penicillin (200단위/ml) 및 streptomycin ($100{\;}{\mu}g/ml$)을 포함한 생리 식염수에 수회 세척하는 살균 조작은 만족할 만한 결과를 가져 왔다. 각종 배지를 사용하였으나 이 중 NCTC 109(Gibco) 또는 NCTC 135(Gibco)에 20% 난환 또는 20% 전란 닥 리액 또는 0.5% yeast를 첨가한 것이 충체 발육 성장에 기초적으로 필요한 것이라 생각되었다. 각종 in vitro 배지 6종류 109개의 시험관에서 시험하였던 바 248마리 충체 중 11마리의 충체가 배양 후 평균 16.1일에 제 3시기에 도달하였으나 더 이상 성장하지는 못하신다. 4. 발육 중인 5일 테지 13일째 된 닭 대아(chick embryo)의 잔뇨막 상에서 $37~38^{\circ}C$로 3일 내지 13일간 배양 한 후 회수한 충체를 관찰한 라 이들의 발육 및 성장이 퀴 또는 병아리에서보다 상당히 지연되기는 하였으나 모든 발육 시기를 볼 수 있었다. 배양 충체에서 34일째는 아무런 발육을 인정할 수 없었으나 봬양 후 평균 5일 내지 10일 사이에 처음으로 제 1 내지 제 2시기 충체가, 평균 제 8일째에서는 제 3시기 충체를 발견할 수 있었다. 한편 소수이기는 하나 산란 시기의 성충 또는 그 이상 성숙한 충체도 배양 후 7일 내지 13일 사이에서 회수할 수 있었다. 따라서 닭 장뇨막 상에서도 성숙 과정의 진행이 가능함을 알 수 있었다. 5. In mitre 배지에서 정상 숙주 또는 장뇨막상으로 이식 실험을 시행하였던 바 in vitro에서 in vivo 이식 실험은 성공하지 못하였으나, in vitro에서 장뇨막으로 및 장뇨막에서 in vivo 숙주로의 실험은 충체의 발육 성장과 관계없이 이식에는 성공하여 생존이 가능하였다. 이상의 결과로 F. seoulensis 피낭유충을 in vitro에서 보존할 수 있을 뿐 아니라, 1~2일 된 갓난 병아리 또 는 닭 장뇨막 상에서 저율이기는 하나 성숙시킬 수 있음을 알 수 있었다.

• 대한한방피부미용학회지
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• 제1권1호
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• pp.53-90
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• 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
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• 제64권6호
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• pp.1105-1116
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• 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
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.302-305
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• 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
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• 제30권11호
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• pp.1785-1791
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• 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.