• Journal of Plant Biotechnology
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• 제47권1호
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• pp.73-77
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• 2020

This study was conducted to examine the vitality of Bletilla striata Rchb.f. seed treated with different concentrations sodium hypochlorite (NaOCl) for different lengths of time. This study also examined the effect of NaOCl treatment times and concentrations on swelling formation and seed germination of B. striata seed. The non-treated B. striata seed had the highest survival rate (82.7%) Treatment with more than 1.5% NaOCl negatively affected the seed survival rate, as compared to concentrations of less than 1%. The swelling formation and seed germination percentages were highest (90.1% and 94.1%, respectively) when seeds were treated with 0.5% NaOCl for 20 min. These results can be used as important basic data for the growth and restoration of B. striata and further suggest the possibility of individual restoration in habitats.

• 한국가금학회지
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• 제41권2호
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• pp.87-92
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• 2014

The avian embryos have been used as a good model to study embryonic development. Due to its unique development in the eggshell, avian embryos can be cultured and hatch in the surrogate eggshell system. In this study, we examined the viability, normal development and hatchability of Korean Oge (KO) chicken embryos in White Leghorn (WL) surrogate eggshells. Donor KO embryos at 3-day and 4-day-old were transferred into recipient WL eggshells, incubated for further 18 days at $37.5^{\circ}C$ with 70% of humidity until hatching. The viability of 3-day-old KO embryos at 7, 14 and 21 day in surrogate eggshell were 70.0%, 43.8% and 23.1%, respectively. In contrast, the viability of 4-day-old KO embryos at 7, 14 and 21 day in surrogate eggshells were 87.1%, 55.6% and 36.0%, respectively. The hatchability of KO embryos transferred into surrogate eggshells at 3-day-old was 23.1%, whereas embryos transferred at 4-day-old was 36.0%. Furthermore, the development of all viable embryos from 3-day group and 4-day group were normal. Our results suggested that culture of KO embryos in WL surrogate eggshells is highly possible, and transfer of donor embryos at 4-day-old may yield higher percentage of hatchability. This study may provide potential knowledge for the conservation of wild and endangered birds through surrogate system.

• Journal of Periodontal and Implant Science
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• 제27권2호
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• pp.329-346
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• 1997

Dental implants have been developed for enhancement of osseointegration. Biocompatibility, bone affinity and surface characteristics of dental implants are very important factors for osseointegration. The aim of the present study was to determine the cytotoxicity and the bone affinity of titanium phosphide(Ti-P) implant material. The Ti-P surface was obtained by vacuum sintering of titanium within compacted hydroxyapatite powder. The composition and the chemical change of the surface were determined by Auger electron spectroscopy. The in vitro cytotoxicity was evaluated by the viability of the bone cells and macrophages obtained from chicken embryo and rat,s peritonium, respectively. For the comparative evaluation, 316L stainless steel, commercially pure titanium and Ti-P materials, prepared in size of 1O.0mm in diameter and 5.0mm in height, were immersed separately in bone cells and macrophages for 10 days. For the evaluation of the in vivo bone affinity, 316L stainless steel, commercially pure titanium and Ti-P materials, prepared in size of 5.0mm in diameter and 10.0mm in length, were implanted after drilling in diameter 5.5mm in femurs of 2 dogs weighing 10Kg more or less. Six weeks after implantation the specimens were prepared for histopathological examination and were observed under light microscope. In comparison of in vitro bone cell viability, Ti-P and commercially pure titanium groups were not significantly different from control group (p>O.1), but 316L stainless steel group was significantly lower than control group(p<0.05). There was no statistical difference in the viability of macrophages between 3 different groups and control group(p>O.l). In comparison of in vivo study, 316L stainless steel and commercially pure titanium showed fibrous encapsulation, but Ti-P showed remarkable new bone formation without any fibrous tissue. The results demonstrate that Ti-P has favorable biocompatibility and bone affinity, and suggest that dental implants with Ti-P surface may enhance osseointegration.

• Journal of Ginseng Research
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• 제45권1호
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• pp.48-57
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• 2021

Background: Ginsenoside Rh2 is well known for many pharmacological activities, such as anticancer, antidiabetes, antiinflammatory, and antiobesity properties. Glycosyltransferases (GTs) are ubiquitous enzymes present in nature and are widely used for the synthesis of oligosaccharides, polysaccharides, glycoconjugates, and novel derivatives. We aimed to synthesize new ginsenosides from Rh2 using the recombinant GT enzyme and investigate its cytotoxicity with diverse cell lines. Methods: We have used a GT gene with 1,224-bp gene sequence cloned from Lactobacillus rhamnosus (LRGT) and then expressed in Escherichia coli BL21 (DE3). The recombinant GT protein was purified and demonstrated to transform Rh2 into two novel ginsenosides, and they were characterized by nuclear magnetic resonance (NMR) techniques and evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide assay. Results: Two novel ginsenosides with an additional glucopyranosyl (6→1) and two additional glucopyranosyl (6→1) linked with the C-3 position of the substrate Rh2 were synthesized, respectively. Cell viability assay in the lung cancer (A549) cell line showed that glucosyl ginsenoside Rh2 inhibited cell viability more potently than ginsenoside Rg3 and Rh2 at a concentration of 10 μM. Furthermore, glucosyl ginsenoside Rh2 did not exhibit any cytotoxic effect in murine macrophage cells (RAW264.7), mouse embryo fibroblasts cells (3T3-L1), and skin cells (B16BL6) at a concentration of 10 μM compared with ginsenoside Rh2 and Rg3. Conclusion: This is the first report on the synthesis of two novel ginsenosides, namely, glucosyl ginsenoside Rh2 and diglucosyl ginsenoside Rh2 from Rh2 by using recombinant GT isolated from L. rhamnosus. Moreover, diglucosyl ginsenoside Rh2 might be a new candidate for treatment of inflammation, obesity, and skin whiting, and especially for anticancer.

• 한국수정란이식학회지
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• 제21권3호
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• pp.263-271
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• 2006

본 연구는 미니 돼지와 일반 돼지(Duroc)의 동결융해 후 정자의 수정 능력을 비교하여 동결 보존체계의 기틀을 확립하고자 하였다. 정액 제조는 수압법으로 정액 채취하여 1차 희석하였다. 동결은 LEY (1차: 11% ${\alpha}$-lactose+egg yolk, 2차: 1차 동결액+glycerol+OEP) 동결액을 이용하여 동결을 실시하여 동결 보존하였다. 동결 정액의 융해는 0.5 ml straw를 각각 20, 37 및 $50^{\circ}C$ water bath에서 1분, 40 초 및 10초간 융해하여 세척 과정 후 BTS 5 ml를 첨가하여 $37^{\circ}C$에서 배양하였다. 정자 성상 검사로는 기형율(Rose Bengal staining), 첨체율(Chlortetra-cycline staining) 및 생존율(SYBR-14/PI staining)등을 배양 후 0, 3, 6, 9 및 12시간에서 검토하여 다음과 같은 결과를 얻었다. 정자의 보존 시간에 따른 기형율은 동결 융해 직후 miniature pig가 $19.5{\pm}1.7%$ 로 Duroc의 $13.9{\pm}0.3%$에 비해 유의적으로 높았다. (p<0.05). 첨체 검사에서 수정능 획득이 일어나지 않은 F pattern은 동결 융해 후 miniature pig와 Duroc 종 정액이 $24.1{\pm}2.8%$와 $37.9{\pm}1.1%$로 Duroc 종에서 유의적(p<0.05)으로 높게 나타났으며, 동결 융해후 miniature pig와 Duroc 종의 AR pattern은 $21.1{\pm}1.6%$ 및 $15.5{\pm}2.2%$로 miniature pig가 유의적(p<0.05)으로 높게 나타났다. 융해 온도별 생존율에서는 20과 $37^{\circ}C$에서는 두 종간에서 유의적 차이는 없었으나, $50^{\circ}C$에서는 miniature pig가 $63.8{\pm}3.6%$로 $47.4{\pm}3.2%$인 Duroc 종에 비해 유의적(p<0.05)으로 높게 나타났다. 본 연구의 결과로부터 첨체율과 기형율에 대한 연구를 보완함으로써 miniature pig정액의 안정적인 동결 체계를 확립할 수 있을 것으로 판단된다.

• 한국수정란이식학회지
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• 제33권3호
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• pp.129-137
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• 2018

Acute vascular rejection has been known as a main barrier occurring in a xenograted tissue of alpha 1,3-galactosyltransferase knock-out (GalT KO) pig into a non-human primate (NHP). Adenosine which is a final metabolite following sequential hydrolysis of nucleotide by ecto-nucleotidases such as CD39 and CD73, act as a regulator of coagulation, and inflammation. Thus xenotransplantation of CD39 and CD73 expressing pig under the GalT KO background could lead to enhanced survival of recipient NHP. We constructed a human CD39 and CD73 expression cassette designed for endothelial cell-specific expression using porcine Icam2 promoter (pIcam2-hCD39/hCD73). We performed isolation of endothelial cells (pAEC) from aorta of 4 week-old GalT KO and membrane cofactor protein expressing pig ($GalT^{-MCP/-MCP}$). We were able to verify that isolated cells were endothelial-like cells using immunofluorescence staining analysis with von Willebrand factor antibody, which is well known as an endothelial maker, and tubal formation assay. To find optimal condition for efficient transfection into pAEC, we performed transfection with GFP expression vector using four programs of nucleofection, M-003, U-023, W-023 and Y-022. We were able find that the program W-023 was optimal for pAEC with regard to viability and transfection efficiency by flow cytometry and fluorescent microscopy analyses. Finally, we were able to obtain $GalT^{-MCP/-MCP}/CD39/CD73$ pAEC expressing CD39 and CD73 at levels of 33.3% and 26.8%, respectively. We suggested that pACE isolated from $GalT^{-MCP/-MCP}$ pig might be provided as a basic resource to understand biochemical and molecular mechanisms of the rejections and as an alternative donor cells to generate $GalT^{-MCP/-MCP}/CD39/CD73$ pig expressing CD39 and CD73 at endothelial cells.

• 한국수정란이식학회지
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• 제22권2호
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• pp.81-87
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• 2007

본 연구에서는 수컷 랫드에 DEHP를 투여하여 실험적으로 생식 독성을 유발하고, vitamin E와 catechin을 단독 및 병용 투여하여 수컷 랫드에서 정소의 조직학적 변화, 정액 특성의 변화 및 정자의 운동성 변화 등을 조사하여 그 예방 효과를 알아보고자 하였다. DEHP를 투여한 군은 대조군에 비해 정자농도, 정자 생존율, 정상 정자율이 감소하였다. DEHP에 의한 생식 독성을 예방하기 위해 vitamin E와 catechin을 단독 또는 병행 투여한 결과, DEHP를 투여한 군에 비하여 정자 농도, 정자 생존율, 정상 정자율이 증가하였다. 이러한 결과를 종합해 볼 때, catechin과 vitamin E의 투여는 DEHP에 의한 생식 독성을 예방하는 효과가 있는 것으로 사료된다.

• 한국수정란이식학회지
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• 제10권1호
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• pp.53-63
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• 1995

The effects of different protein sources (serum vs bovine serum albumin), growth factors (EGF and PDGF) and co-culture with various type of somatic cel1s (BOEC, MEF and BRL) on the in vitro development of in vitro matured / in vitro fertilized bovine oocytes were examined, and the viability of frozen/thawed embryos derived from IVM /IVF was examined. Cell numbers of blastocysts were also counted. In Experiment 1, CR$_1$aa with serum was superior to CR$_1$aa with BSA in producing morulae plus blastocysts from IVM /IVF oocytes(24.4% vs 30.4%, p>0.05). In Experiment 2, more morulae plus blastocysts(42.3%) were produced in CR$_1$aa containing long /ml EGF than in the control CR$_1$aa(33.3%). In Experiment 3, 2- to 8-cell embryos derived from IVM /IVF oocytes were randomly allotted to one of 4 culture groups : a) CR$_1$aa ; b) CR$_1$aa + ing /ml PDGF ; CR$_1$aa + Sng /ml PDGF ; CR$_1$aa + lOng /ml PDGF ; culture resulted in 21.3, 51.2, 41.4 and 45.9%(p<0.05), respectively, developing into morulae and blastocysts. In Experiment 4, 0 and Sng /ml PDGF added to CR$_1$aa coculture with BRL or BOEC yielded 47.5, 42.5, 33.8 and 41.6% morulae and blastocysts, respectively. In Experiment 5, the proportion of embryos into morulae and blastocysts was highest in CR$_1$aa with MEF coculture group(50.9%) compared to any other group(CR$_1$aa, 22.3%; CR$_1$aa+BRL, 32.9%; CR$_1$aa+BOEC, 33.8%, p>0.05). In Experiment 6, survival rate of blastocysts produced by in vitro fertilization when cryoprotectant was removed in 0.7M glycerol+0.7M sucrose and 0.7M sucrose solution for 10 min. after thawing at 2$0^{\circ}C$ (Exp. H, 58.8%) was slightly higher than when cryoprotectant was removed 10%, 6.7% and 3.3% glycerol for 10 min. after thawing at 37$^{\circ}C$ (Exp. I, 54.3%). These study indicate that growth factors and somatic cell co-culture can increase the proportion of embryos that develop into morulae and blastocysts without an increase in the cell number and frozen /thawed method employed this experiment was not different.

• 한국수정란이식학회지
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• 제21권4호
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• pp.339-344
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• 2006

본 연구는 염소 액상 정액의 활용에 적합한 희석액을 검토하여, 국내 염소 인공수정 실용화를 위한 기초 자료를 확보하기 위하여 실시되었다. 두 마리의 흑염소 수컷으로부터 전기 자극법으로 정액을 채취하였다. 채취된 정액은 BTS로 희석하여 500g에서 5분간 원심분리하여 정장을 제거하고, BTS, Modena, Triladyl$^{(R)}$로 희석하여 $4^{\circ}C$ 냉장고나 $17^{\circ}C$ 온장고에서 8일간 보관되었다. BTS와 Modena로 희석된 정자의 운동성은 하루 만에 소멸되었으며, BTS로 희석된 정액을 $17^{\circ}C$에 보관하였을 때 2일째에 30$\sim$45%로 떨어졌으며, Modena에서는 3일째까지 인공수정에 가능한 활력을 유지하였다. 한편 Triladyl$^{(R)}$로 희석하여 $4^{\circ}C$나 $17^{\circ}C$에서 보관한 경우에는 좀더 좋은 생존성을 나타내었으며, 4일까지도 인공수정이 가능한 운동성을 보여주었다.

• 한국수정란이식학회지
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• 제29권2호
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• pp.163-169
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• 2014

The objective of this study was to evaluate the characteristics of Korean Native Cattle sperm frozen-thawed with L-cysteine and/or catalase. The semen from bulls was collected by the artificial vagina method, and Triladyl containing 20% egg-yolk and/or L-cysteine (L), catalase (C) and L-cysteine + catalase was added to the diluted semen for cryopreservation. The results showed that sperm viability was significantly higher in the L-cysteine + catalase ($69.49{\pm}3.16%$) group than in the control ($60.5{\pm}3.94%$) group (p<0.05). Acrosome damage was significantly lower in the L-cysteine ($17.12{\pm}1.08%$) group than in the control ($21.46{\pm}1.14%$), catalase ($20.54{\pm}0.76%$), and L-cysteine + catalase ($19.29{\pm}0.65%$) groups (p<0.05). In addition, the level of intact mitochondria in the spermatozoa was significantly higher in the L-cysteine ($58.65{\pm}1.39%$) group than in the control ($50.63{\pm}2.37%$) group (p<0.05). The hydrogen peroxide level in the frozen-thawed sperm was significantly lower in the L-cysteine ($3.74{\pm}1.66%$), catalase ($4.65{\pm}1.87%$), and L-cysteine + catalase ($8.11{\pm}2.15%$) groups than in the control ($13.22{\pm}1.6%$) group (p<0.05). The glutathione level was significantly higher in the L-cysteine ($1.33{\pm}0.03%$) group than in the control ($1.08{\pm}0.06%$), catalase ($1.05{\pm}0.02%$) and L-cysteine + catalase ($1.11{\pm}0.03%$) groups (p<0.05). In conclusion, L-cysteine and catalase could protect the membrane of Korean Native Cattle sperm from damage during sperm cryopreservation. Especially, L-cysteine was more effective for keeping acrosomes and mitochondria intactness during sperm cryopreservation.