• Clinical and Experimental Reproductive Medicine
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• v.28 no.1
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• pp.33-40
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• 2001

Objective: This study was to establish the human embryonic stem (ES) cells derived from frozen-thawed blastocyst stage embryo that were destined to be discarded after five years in routine human IVF-ET program. Methods: Frozen-thawed and survived human blastocysts were treated by immunosurgery, and recovered ICM cells were cultured onto STO feeder cell layer and ICM colony was subcultured by mechanical dissociation into clumps. To identify ES cell, alkaline phosphatase staining and expression of Oct4 in replated ICM colonies were examined. Also, to examine the possibility of ES cell differentiation, retinoic acid (RA), basic fibroblast growth factor (b-FGF), nerve growth factor (NGF) were added in culture medium. In addition, to classify the specific cell type, differentiated cells were stained by indirect immunocytochemistry. Results: One ICM colony recovered from frozen-thawed six blastocysts was subcultured, continuously replated during 40 passage culture duration without differentiation. Subcultured colonies were strong positively stained by alkaline phophatase. When the expression of Oct4 in cultured ES colony was examined, Oct4b type is more clearly indicated than Oct4a one although there was not detected in embryoid body or differentiated cells. In differentiated cardiomyocytes from ES colony, cells were beaten regularly (60 times/min). In differentiated neural cells from ES colony, neurofilament (NF) 200 kDa protein, microtubule associated protein (MAP) 2 and ${\beta}$-tubulin of specific marker in neurons, glial fibrillary acidic protein (GFAP) of specific marker in astrocytes and galactocelebrocide (GalC) of specific marker in oligodendrocytes were confirmed by indirect immunocytochemistry. Also, muscle cells were detected by indirect immunocytochemistry. In addition, ES colonies can be successfully cryopreserved. Conclusion: This study suggested that establishment of human ES cells can be successfully derived from frozen-thawed blastocysts that were destined to be discarded, and obtained specific cell types (cardiomyocytes, neurons and muscle cells) through the in vitro differentiation procedures of ES cells.

• Journal of The Korean Society of Grassland and Forage Science
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• v.35 no.4
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• pp.297-302
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• 2015

This experiment was conducted to evaluate the agronomic characteristics and forage production of Italian ryegrass cultivars at three locations (Gurae, Sungju and Cheonan) from 2013 to 2014. The experiment employed a randomized complete block design with three replications. The seven cultivars used in this experiment were 'Korwinearly', 'Florida80', 'YounbongI', 'YounbongII', 'YounbongIII', 'KeimyungI' and 'KeimyungII' varieties. Italian ryegrass varieties were seeded on October 4 at Gurae, October 8 at Seongju and October 3 at Cheonan in 2013. The Italian ryegrass varieties were harvested on April 26 at Gurae and Seongju, and on April 24 at Cheonan in 2014. Although the seven cultivars had resistance to foliar diseases and insects and high winter survival, they has little lodging resistance. The heading dates for 'Younbong I', 'Younbong II' and 'Keimyung I' were earlier than for other varieties at the three locations. Dry matter (DM) content and plant height in the early maturing cultivars were higher than in other cultivars at the three locations. The fresh and DM yields of the early maturing cultivars were also higher than others at the three locations. The results of this experiment indicate that the heading date for early maturing cultivars is earlier, and the plant height is higher than for other cultivars. Early maturing cultivars had higher DM content as well as fresh and DM yields of Italian ryegrass. Therefore, early maturing cultivars should be recommended as high-yield forage in double-cropping systems because of their superior production of forage and early heading date.

• Journal of The Korean Society of Grassland and Forage Science
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• v.34 no.2
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• pp.81-86
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• 2014

This study was conducted to evaluate the effect of different seeding dates on agronomic characteristics, forage quality, dry matter (DM) and dry matter digestible (DDM) yields of sudangrass hybrid (Sorghum bicolor(L.) Moench) and oat (Avena sativaL.) at Seongju in Kyeongbuk from 2012 to 2013. The experiment was arranged in randomized complete block design with three replications. Sudangrass was seeded five times seeding dates of 10 days interval ranging from $1^{th}$ August to $10^{th}$ September in 2012 and 2013. Oat was seeded three times seeding dates of 10 days interval ranging from $20^{th}$ August to $10^{th}$ September in 2013. Sudangrass was harvested on $26^{th}$ October, 2012 and $2^{nd}$ November, 2013. Plant length and DM content decreased with delaying seeing date. The DM and DDM yields of sudangrass were higher (P<0.05) in first seeding date than all other seeding dates and decreased with delaying seeding dates (P<0.05). The DM and DDM yields of oat were higher (P<0.05) in first and second seeding dates than third seeding date. The results of this study indicated that early seeding of sudangrass would be a recommended than late seeding after $20^{th}$ of August, and late seeding of oat after $30^{th}$ August would be recommened for DM and DDM yields in the Southern Korea.

• Journal of The Korean Society of Grassland and Forage Science
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• v.33 no.2
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• pp.117-122
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• 2013

This experiment was conducted to evaluate agronomic characteristics, forage quality, and dry matter (DM) yield of hairy vetch 'Cold green', common vetch 'Maxivesa', 'Wild common', 'Rasina', 'Morava', and 'Blanchefleur', at Seongju in Kyeongbuk from 2010 to 2012. The experiment was arranged in randomized complete block design with three replications. Vetchs were seeded on the 10th of Oct. 2010 and on the 3rd of Oct. 2011. Vetch plants were harvested on the 4th of May 2011 and on the 10th of May in 2012. 'Cold green' and 'Wild common' exhibited greater cold tolerance than other cultivars. Fifty percent-flowering of 'Cold green' occurred on the 22nd of Apr., whereas that of 'Wild common' occurred on the 26th of Apr. and those of 'Rasina' and 'Blanchefleur' occurred on 28th of Apr. and that of 'Maxivesa' occurred on 17th of May. The DM yield of 'Cold green' was higher (p<0.05) than all other cultivars, whereas the DM yield of 'Wild common' was higher (p<0.05) than those of the remaining 4 cultivars. According to the results obtained from this study, it is suggested 'Wild common' vetch is be recommendable for cultivation where increased DM production is sought.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.2
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• pp.129-134
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• 1998

The objective of this study was to test whether the developmental capacity of human multi-pronuclear (PN) zygotes after ultra-rapid freezing using EM grid can be maintained. For this experiment, multi-PN zygotes which produced in human IVF program were used as an alternatives of normal 2PN zygotes, and they were separated into 3PN or $\geq4PN$ zygotes to compare their in vitro development and cryoinjury according to PN number. As freezing solution, EFS30 which consisted of 30% ethylene glycol, 18% bcoll, 0.5 M sucrose and 10% FBS added D-PBS was used. The result obtained in this experiment was summarized as follows; When the multi..PN zygotes were ultrarapidly frozen and thawed, the high mean percentages (85.5%) were survived. Also when the cleavage rates between control and freezing group were compared with PN number, there were not significantly different in each group (3PN; 81.3% & 85.4% and $\geq4PN$; 90.0% & 95.7%). When the in vitro development rates after thawing were examined, freezing 3PN group (22.0%) was not differed to control 3PN group (38.5%), although the development result of freezing $\geq4PN$ group (45%) was significantly lower than that of control $\geq4PN$ group (44.4%) (p<0.05). These results demonstrate that developmental capacity of human zygote can be obtained by ultra-rapid freezing method using EM grid and EFS30.

• Korean Journal of Veterinary Research
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• v.38 no.1
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• pp.43-52
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• 1998

서방형 돼지성장호르몬(sustained-release formulation of porcine somatotropin, PST-SR)을 1주 간격으로 6차례 피하 및 근육주사하고 혈액과 조직중의 돼지 성장호르몬(PST)과 insulin-like growth factor 1(IGF-1)의 농도를 측정하여 다음과 같은 결과를 얻었다. 대조군의 혈중 PST와 IGF-1의 농도는 각각 2.41과 95.2 ng/ml 이었다. 1. PST-SR을 투여한 후 PST의 혈중농도는 8시간만에 최대에 도달하여(30 ng/ml) 곧 감소하였다. 혈중농도 반감기(decay half life)는 91~227시간이었다. IGF-1의 혈중농도는 투여후 12시간에 최대에 도달하였으며(165 ng/ml), 이후 서서히 감소되었고 반감기는 77~99시간이었다. 2. 혈중 PST농도-시간의 자료는 제재에서 PST가 유리되는 과정에는 두단계 즉, 투여후 24시간까지의 유리속도가 빠른 단계와 그 이후의 유리속도가 느린 단계가 있음을 보여주었다. 3. 여섯번의 반복투여기간에는 PST의 혈중농도는 투여직후 증가하여 24시간 이후 다음 투여전까지 지속적으로 감소되는 패턴이 반복되었고, 최종투여후 1주일경에는 정상수준으로 회복되었다. 반면에 투여가 반복됨에 따라 매 투여직후의 PST의 혈중 최고치는 다소 증가되는 경향을 보였다(20~40 ng/ml). IGF-1의 혈중농도는 투여가 반복됨에 따라 누적적인 증가현상이 뚜렷하였으며, 이후 2주일후 까지도 정상농도보다 높게 유지되었다(200ng/ml). 임상용량 투여군에서 PST 및 IGF-1의 혈중농도는 투여경로에 따른 차이는 나타나지 않았다. 4. 최종(6번째) 투여후 6, 8, 10, 14일에 조사한 간장, 신장, 소장, 근육, 지방 및 주사부위의 조직중의 PST 농도는 6일째에 이미 대조군 수준으로 회복되었다. IGF-1의 경우 최종투여후 6일에는 간장, 신장, 소장, 지방조직에서 정상보다 높은 농도로 잔류하나 이후 14일까지 모두 대조군 수준으로 감소되었다. 5. 이상의 결과는 본 실험에서 사용된 서방형 PST제제는 최소 1주간 유효성이 유지되며, 동시에 PST는 투여 6일째에, IGF-1은 투여후 14일에 정상수준으로 회복됨을 보여주고 있다.

• Korean Journal of Food Science and Technology
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• v.36 no.5
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• pp.779-783
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• 2004

New food ingredient was developed to eradicate and protect against re-infection of Helicobacter pylori in fermentation broth of lactic acid bacteria (LAB) showing antimicrobial activity against pathogenic microorganisms such as H. pylori and Listeria monocytogenes. LAB strain CBT SL4 was identified as Pediococcus pentosaceus by 16S rDNA sequencing and its culture broth showed antimicrobial activity of 800 AU/mL against H. pylori in optimized fermentation process. Using thin layer concentration system and spray-typed fluid bed drier system, concentrated powder product showing activity of 12,800 AU/g was harvested. Product showed eradication and protection activities against H. pylori infection on feeding test (50 AU/day) using Mongolian gerbil infection model. After 4 weeks therapy of 8,000 AU/day, ${\Delta}13CO_2$ level (DOB30) decreased about 40% in urea breath test on patient with H. pylori infection. Result show concentrated culture product of P. pentosaceus CBT SL4 has eradicating and protecting activities against H. pylori infection and can be used as food-active ingredient for prevention of gastric and duodenum ulcer caused by H. pylori.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.2
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• pp.129-138
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• 2002

Objective: This study was to compare the characteristics between parthenogenetic mES (P-mES) cells and in vitro fertilization mES cells. Materials and Methods: Mouse oocytes were recovered from superovulated 4 wks hybrid F1 (C57BL/6xCBA/N) female mice. For parthenogenetic activation, oocytes were treated with 7% ethanol for 5 min and $5{\mu}g$/ml cytochalasin-B for 4 h. For IVF, oocytes were inseminated with epididymal sperm of hybrid F1 male mice ($1{times}10^6/ml$). IVF and parthenogenetic embryos were cultured in M16 medium for 4 days. Cell number count of blastocysts in those two groups was taken by differential labelling using propidium iodide (red) and bisbenzimide (blue). To establish ES cells, b1astocysts in IVF and parthenogenetic groups were treated by immunosurgery and recovered inner cell mass (ICM) cells were cultured in LIF added ES culture medium. To identify ES cells, the surface markers alkaline phosphatase, SSEA-1, 3,4 and Oct4 staining were examined in rep1ated ICM colonies. Chromosome numbers in P-mES and mES were checked. Also, in vitro differentiation potential of P-mES and mES was examined. Results: Although the cleavage rate (${\geq}$2-cell) was not different between IVF (76.3%) and parthenogenetic group (67.0%), in vitro development rate was significantly low in parthenogenetic group (24.0%) than IVF group (68.4%) (p<0.05). Cell number count of ICM and total cell in parthenogenetic b1astocysts ($9.6{\pm}3.1,\;35.1{\pm}5.2$) were signficantly lower than those of IVF blastocysts ($19.5{\pm}4.7,\;63.2{\pm}13.0$) (p<0.05). Through the serial treatment procedure such as immunosurgery, plating of ICM and colony formation, two ICM colonies in IVF group (mES, 10.0%) and three ICM colonies (P-mES, 42.9%) in parthenogenetic group were able to culture for extended duration (25 and 20 passages, respectively). Using surface markers, alkaline phosphatase, SSEA-l and Oct4 in P-mES and mES colony were positively stained. The number of chromosome was normal in ES colony from two groups. Also, in vitro neural and cardiac cell differentiation derived from mES or P-mES cells was confirmed. Conclusion: This study suggested that P-mES cells can be successfully established and that those cell lines have similar characteristics to mES cells.

• Journal of The Korean Society of Grassland and Forage Science
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• v.32 no.3
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• pp.229-236
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• 2012

This experiment was conducted to evaluate agronomic characteristics and forage productivity of italian ryegrass cultivars at Seongju in the Kyeongbuk from 2010 to 2012. The experiment was arranged in randomized complete block design with three replications. Italian ryegrass cultivars were seeded on Oct. 12 in 2010 and Oct. 3 in 2011. The early-medium maturing cultivars were harvested on 29 Apr. and the medium late ones were harvested on 4 May in 2011. In 2012, the early-medium cultivars were harvested on 28 Apr. and medium late ones were harvested on 12 May. DM (dry matter) yield of the early-medium maturing cultivars 'Yeonbong3', 'Seongnong', 'Winter Hawk', 'Pride' and 'Kowinnearly' was 9,850, 9,778, 9,486, 9,363 and 9,267 kg/ha and DM yield of those were significantly higher (p<0.05) than other seven cultivars or ones. DM yield of the medium-late maturing cultivars 'Tetragold', 'Hwasan 101', 'Jumbo', 'Sungrazer', 'Master', 'SelectIV' and 'KB Royal' was 9,542, 9,492, 9,103, 8,981, 8,903, 8,870 and 8,681 kg/ha and DM yield of those were higher (p<0.05) than other three. Cold tolerance for 'Hwasan 101' was higher than others, in medium-late maturing cultivars. According to the results obtained from this study, it is suggested that early-medium maturing cultivars would be recommendable for DM production.

• Korean Journal of Food Science and Technology
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• v.10 no.3
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• pp.313-319
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• 1978

In order to investigate the effect of dietary long chain fatty acids on fatty acid biosynthesis of liver in birds, single comb White Leghron male chicks were fed a fat-free diet an diets containing margaric, stearic and linoleic acids and liver lipid components and liver and plasma fatty acid distributions were compared. Total lipids of tissues were extracted with a chloroform-methanol mixture. The lipid components were determined by thin layer chromatography and fatty acid distribution of lipid fractions were determined by gas liquid chromatography. Fatty acid feeding did not affect liver lipid components. When margaric acid(17 : 0), was fed, 17:0 and heptadecenoic acid(17:1) appeared in every lipid fractions of liver and plasma, and distribution values of these acids were not significantly different between the lipid fractions of liver. In blood plasma of the 17 : 0 fed chicks, however, significantly higher distribution values of 17 : 0 and 17.1 were observed in the triglyceride fraction and in the cholesterol ester fraction, respectively. Dietary stearic acid (18 : 0) did not show any effect on the distribution of 18 : 0 in every lipid fractions of liver but showed a significantly higher distribution value of 18 : 0 in the free fatty acid fraction of plasma. When linoleic acid (18 : 2) was fed, every lipid fractions of liver and plasma contained 18 : 2, especially a significantly higher distribution value was observed in the phospholipid fraction of liver. Dietary margaric and linoleic acids tended to decrease the distribution value of endogenously synthesized palmitoleic (16 : 1) and oleic (18 : 1) acids in liver.