• 한국가축번식학회지
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• 제20권2호
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• pp.191-196
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• 1996

The objective of this study were to investigate the effects of culture media and additives on the development of bovine in vitro matured(IVM) and in vitro fertilized(IVF) oocytes. In experiment 1, bovine oocytes were cultured in droplets of TC 199 supplemented with 10% fetal bovine serum(FBS) with or without hormones (5$\mu\textrm{g}$/ml FSH, 5$\mu\textrm{g}$/ml LH, 1$\mu\textrm{g}$/ml E2). Cleavage rates of embryos cultured for 40~44hrs after IVF were higher when embryos were cultured in TC 199 supplemented hormones (68.1%, 921/35) than without hormones (52.7%, 77/146), but the percentages of development beyond morulae stage were not difference (20.7%, 19.4%). In experiment 2, the effects of various media such as TC 199, synthetic oviduct fluid(SOF), CR1aa with different energy source (fatal bovine serum, FBS; bovine serum albumin, BSA) on developmental capacity of IVM/IVF bovine embryos were investigated. The developmental rates into morulae and blastocysts were 27.1, 10.7, 6.3 and 0%, respecitvely, in CR1aa plus 3mg/ml BSA, SOF plus 10% FBS, TC 199 plus 10% FBS, SOF plus 3mg/ml BSA. In experiment 3, the comparisons of bovine embryos developed to morulae and blastocysts in different culture media (TC 199, SOF, CR1aa, Menezo's B2) were investigated. The developmental capacity beyond morulae stage were 32.9, 26.6, 11.1 and 7.1%, respectively, in Menezo's B2 plus BSA, CR1aa plus BSA, SOF plus BSA, TC 199 plus FBS medium. The cell numbers of the blastocyst were not different in different cultrue media. In experiment 4, bovine embryos were co-cultured with vobine oviduct epithelial cells(BOEC) in TC 199 plus FBS, SOF plus BSA, CR1aa plus BSA, Menezo's B2 plus BSA. The morula and blastocyst rates were 44.7, 32.9, 26.0 and 23.3%, respectively, in CR1aa TC 199, SOF, and Menezo's B2 medium. The cell numbers of the blastocyst were similar to those of blastocyst developed in different culture media.

• The Plant Pathology Journal
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• 제21권3호
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• pp.262-269
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• 2005

An antiviral producing bacterial strain was isolated from a ginseng rhizosphere in Kangwon province of Republic of Korea. In order to identify the bacterial strain, microbiological, physiological and biochemical tests were performed, along with RAPD, 16S rRNA, 16S-23S rRNA ITS (intergenic spacer region) and DNA-DNA hybridization analyses. The bacterium was found to be a strain of Pseudomonas fluorescens, which was designated as Gpf01. The strain was grown in Muller-Hinton (MH) broth, and the culture supernatant obtained was filtered through a $0.45{\mu}l$ filter. It was further boiled at $100^{\circ}C$ and tested in two experiments for its ability to control a yellow strain of Cucumber mosaic virus (CMV-Y). In the first experiment, boiled culture filtrate (RCF) was treated on one half of the leaves of Chenopodium amaranticolor followed by CMV- Y inoculation on both halves. In the second experiment, BCF was treated on the lower leaves of Nicotiana tobacum var. Xanthi-nc, with the CMV-Y mechanically inoculated onto the upper untreated leaves. In the first experiment, BCF treatment was able to considerably reduce the number of viral lesion, and in the second experiment, plants treated with BCF showed no visible viral symptoms compared to the Muller-Hinton (MH) media treated controls 15 days post inoculation (dpi), and remained symptomless throughout the study period. Thus, Gpf01, identified as P. fluorescence, was able to produce an antiviral component in the culture filtrate, which was found to be heat stable, non-phytotoxic and effective in local as well as systemic hosts of CMV.

• 한국연초학회지
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• 제20권2호
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• pp.172-177
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• 1998

The culture filtrate of Streptomyces sp. B25, which was identified in this experiment, was tested for the control of tobacco mosaic virus (TMV) with the susceptible tobacco cultivar, NC 82, under the field conditions following the preliminary examination of its characters for TMV control. Control efficacy of the culture filtrate against TMV infection continued over 50% up to 6 days after treatment, and its systemic effect was about 30% of the direct effect. In field conditions control efficacy of the culture filtrate against TMV infection was 95.3 % at 2 weeks after TMV inoculation, and decreased to 58.3 % at 3 weeks after inoculation. Five fold-dilution of the culture filtrate showed about half of the control efficacy by the stock culture filtrate. Analysis of the antibiotic material responsible for the inhibition of TMV infection through nuclear magnetic resonance experiments revealed that the antibiotic is antimycin $A_1$, which is firstly reported as an anti-phytoviral antibiotic in this experiment.

• Reproductive and Developmental Biology
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• 제34권3호
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• pp.207-211
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• 2010

Current developments in IVF and animal cloning have resulted in increasing demand for large quantities of oocytes and ovarian follicles at specific stages of development. These medical and scientific needs may be met by developing an optimal culture system for preantral follicles. In this study, we investigated the growth of porcine preantral follicle cultures in different media and in the presence and absence of serum. Follicles were manually dissected from ovaries obtained from prepubertal gilts at a local slaughterhouse, and cultured for 3 days in M199 or NCSU23 medium supplemented with porcine FSH, transferrin, L-ascorbic acid and insulin. Follicle diameters were measured on day 1 and 3 of culture. In Experiment 1, the effect of supplementing culture medium with fetal calf serum (FCS) on porcine preantral follicle growth was examined. In the group of cultures supplemented with FCS, follicle diameter after 3 days of culture, survival rate and antrum formation rate in the FCS group were significantly higher than those of the control group. In Experiment 2, the effects of culture medium (M199 and NCSU23) on follicle growth were compared. Follicle diameters were increased in the M199 group, compared with those in NCSU23 (p<0.05), but we observed no significant differences in survival and antrum formation rates between cultures grown in the two media. In conclusion, supplementation of the culture medium with serum enhances preantral follicle growth and antrum formation, and M199 is superior to NUSU23 for porcine preantral follicle culture in vitro.

• 한국수정란이식학회지
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• 제28권1호
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• pp.41-48
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• 2013

This study examined the motility of either the unattached(upper) or attached(lower) Hanwoo sperm to bovine oviduct epithelial cell(BOEC) monolayers to determine whether there are any changes in their motility during co-culture. The cleavage and blastocyst development rate were compared among different preincubation methods in-vitro, after oocytes were fertilized in-vitro with Hanwoo sperm on BOEC monolayers. The motility of frozen-thawed sperm in BOEC co-culture group was significantly higher than controls, especially at 5 hours and 6 hours (p<0.05) of incubation, in sperm treatment medium without heparin and caffeine. The motility of frozen-thawed sperm in BOEC co-culture group was significantly higher than controls, especially at 3 hours (p<0.05) and 6 hours (p<0.01), in sperm treatment medium containing heparin and caffeine. The motility of the attached( lower) sperm was significantly higher than the unattached(upper) sperm during co-culture with BOEC at all times(p<0.01 or p<0.05), except for 6 hours. After Hanwoo oocytes were fertilized in-vitro with the sperm that had been co-cultured with BOEC in sperm treatment medium containing heparin and caffeine, we determined the cleavage and blastocyst development rate, according to the preincubation methods. Both the cleavage and blastocyst development rate from 2 hour preincubation group were the highest, but significant difference was not recognized. These results show that BOEC plays an important role on sperm hyperactivation related to capacitation regardless of heparin and caffeine in sperm treatment medium. However, oviduct epithelial cell had no significant effect on the development of embryos after in-vitro fertilization in the presence of added heparin and caffeine in sperm treatment medium.

• 한국동물생명공학회지
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• 제38권2호
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• pp.70-76
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• 2023

Background: Despite considerable technological advancements, polyspermy remains a significant challenge in in vitro fertilization (IVF) procedures in pigs, disrupting normal embryonic development. Here, we aimed to determine whether optimal fertilization conditions reduce the polyspermy incidence in pigs. Methods: In vitro-matured oocytes were co-incubated with sperm according to a modified two-step culture system. Results: In the first experiment, oocytes were briefly co-incubated with sperm, washed in IVF medium, and then moved to fresh IVF medium for 5 or 6 h. Although the 6 h sperm-free cultured group had a higher penetration rate than the 5 h cultured group, the polyspermy rate significantly increased in the 6 h sperm-free cultured group. The gamete co-incubation period was either 20 or 40 min. The 40 min cultured group had a higher rate of blastocyst formation and number of total cells in blastocysts than the 20 min cultured group. In experiment 2, oocytes were inseminated with sperm separated by Pecroll treatment. Percoll treatment increased the rate of oocyte penetration and blastocyst formation compared to the control. In experiment 3, fertilized oocytes were cultured in 25 µL microdroplets (10 gametes/drop) or 500 µL (100 gametes/well) of culture medium in 4-well plates. The large volume of medium significantly reduced the number of dead oocytes and increased the rate of blastocyst formation compared to the small volume. Conclusions: Collectively, these results demonstrate that various fertilization conditions, including modified co-culture period, active sperm separation, and culture medium volume, enhance fertilization efficiency and subsequent embryonic development by decreasing polyspermy occurrence.

• 한국동물학회지
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• 제37권2호
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• pp.255-261
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• 1994

Recombinant human pBrathyriod hormone related peptide (1-341 (rhPTHrP) has been known to stimulate bone resorption in intact bone tissue culture system. Osteoclast has been known as a primary responsible cell for bone resorption. To examine the effect of rhPTHrP on this cell, we employed disaggregated rat osteodast culture. As a result, we found that rhPTHrP sisnificBntly elevates both the number and total area of resorbed pits in this culture. On the other hand, the conflicting results between disagsregated rat osteoc13st culture and Ca2+-deficient hen osteoclast culture system have been a big obstacle for the progress of bone research. To verify the differences between rat 3nd chick osteoclast system, we performed the same experiment using chick embryonic osteoclast. Since the similar results were obtained from the disaggregated chick osteoclast culture, the discrepancy between chick and rat osteoclast culture study seemed to be due to the difference in culture method, rather due to the species-difference.

• 한국수정란이식학회지
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• 제10권3호
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• pp.229-236
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• 1995

The experiments reported here take advantage of the large number of in vitro matured and in vitro fertilized(IVM /IVF) bovine oocytes which can be produced, permitting the design of controlled experiments to establish a simple defined medium for the study of early embryo requirements. A total of 1,386 IVM /IVF oocytes were used to compare a simple defined medium(KSOM) with more complex culture conditions used successfully for culture of bovine embryos but do not permit study of specific requirements. All experiments were extensively replicated factorials. In Experiment 1, KSOM was superior to Menezo B$_2$ medium in producing morulae plus blastocysts from IVM /IVF oocytes(33 vs 20%, P<0.()5). The yield of morulae plus blastocysts with KSOM was 22% and with BRLC added was 30%. In Experiment 2, (a 2x2 factorial of KSOM with or without BRLC and 0, 1 ng /ml of platelet derived growth factor, PDGF) more morulae plus blastocysts (40%) were produced in KSOM-BRLC co-culture containing 1 ng /ml PDGF than in the control KSOM(12%). In Experiment 3, there was no dose response when 0, 1 and 5 ng /ml of PDGF were added. The results with simple defined KSOM medium are sufficiently promising to indicate that specific requirements of the embryo may be examined in future studies with KSOM as a base.

• 한국작물학회지
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• 제46권1호
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• pp.47-52
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• 2001

In order to develop more stable seedling stand method in wet direct rice seeding culture, the effect of making the drainage ditches was studied in both methods of broadcast seeding on floody paddy surface and puddled-soil drill seeding. In a broadcast seeding on floody paddy surface, the ditching after seeding using a tractor or small ditch maker improved the seedling stand, and reduced the floating seedling and herbicide injury by accelerating the drainage. Suitable ditching time was at 2 days after seeding (DAS) for the tractor and at 3 DAS for the ditch maker. In the puddled-soil drill rice seeding culture, the ditching before seeding with a ditch maker at 3 days after draining effectively improved the seedling stand due to quick draining through well formed the seeding furrows. In the puddled-soil drill seeding, seedling stand number showed higher in both ditching plots synchronized with seeding compared with both only draining treatment at 1 DAS without ditching and the flooding plot condition for 6 DAS. And the suitable ditching depth was 6 cm, as considered the ditching status and drained status. These results suggest that the ditching in wet direct rice seeding is an resonable practice for improving the seedling stand through the accelerating drainage of field.

• 한국작물학회지
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• 제37권5호
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• pp.419-424
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• 1992

콩에서의 조직배양 가능성을 알아보고자 콩 유묘를 기내에서 배양시킨 뒤, 여러 부위를 채취하여 각기 다른 배지 치상한 결과는 다음과 같다. 1. 줄기정단조직 배양에서는 대부분 shoot가 분화되어 완전한 식물체를 얻을 수 있었다. 2. 자엽절 조직배양에서는 multiple shoots가 나타나 대량증식에 유용했으나 배지에 따른 영향이 컸다. 3. 배축 및 상배축 조직배양에서는 callus만 분화되엇다. 4. 엽조직배양에서는 callus 분화 후 계대배양했을 때 shoot가 분화되었으나 완전한 식물체를 얻기는 힘들었다.