• Clinical and Experimental Reproductive Medicine
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• v.25 no.2
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• pp.109-113
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• 1998

Objective: To ananlyze the direct effect of nitric oxide (NO), generated from sodium prusside (SNP) on the embryo developments in reproductive process. Design: Ova from mouse were treated to allow fertilization in in vitro culture. And the samples of fertilized ova were alloted into five alliqutos. Each alliquot was cultured in media treated with either concentration at 0 (n=92), $25{\mu}M$ (n=84), $50{\mu}M$ (n=80), $100{\mu}M$ (n=77), $500{\mu}M$ (n=54) of SNP. Main Outcome Measure: Rates of embryonal cell cleavages, viability and cell morphology were assessed during in vitro fertilization and culture. Results: As analyse the cell cleavage at 24 hours after in vitro culture of fertilised egg in variuos NO concentration, all of egg cells of each alliquot were developed to $2\sim4$ cell stage. But the alliquot of egg cells treated with $50{\mu}M$, which were totally degenerated. And also all embryonal cells of each alliquot were developed to 8 cell stage and morula stage on culture continuosly. And the embryonal cells of each alliquot were analysed at 24 and 48 hours following the in vitro culture. The rates of cell fragmentation and fusion were $4.2{\pm}3.4%$ in control group which is not treated with NO, while experimental groups was high, as rated $23.4{\pm}6.2%$ in $25{\mu}M$, $28.2{\pm}5.7%$ in $50{\mu}M$ and $32.1{\pm}6.4%$ in $100{\mu}M$ concentration of NO. Accordingly the rate of abnormal morphology of embryonal cell in control was lower significantly than that in each alliquot of experimental groups (p<0.05). And the degenerated rates of embryonal cells were 0% in control, $17.8{\pm}6.7%$ in $25{\mu}M$, $23.6{\pm}4.7%$ in $50{\mu}M$ and $26.8{\pm}11.2%$ in $100{\mu}M$ at 8 cells and morula on culture of 48 and 72 hours. On the examination of embryonal cells developed to blastocyst through in vitro culture, the rates of degenerated cells were $16.8{\pm}7.2%$ in control, $37.5{\pm}6.2%$ in $25{\mu}M$, $73.4{\pm}4.6%$ in $50{\mu}M$, 100% in $100{\mu}M$. Conclusion: This results suggeted that the NO in any concentrations is harmful on embryos in view of morphology as well as viability of cell, and the toxicity of NO on embryo is stronger at condition in higher concentration of NO.

• Korean Journal of Animal Reproduction
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• v.16 no.3
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• pp.209-215
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• 1992

These studies were carried out to overcome 2-cell block and in vitro development to blastocysts in vitro fertilization of mouse embryos. The unfertilized ova were obtained by superovulation in ICR mice of 4 to 6 weeks old. Tyrode's 280 solution was used as basal media, and the pH range of media examined was designed from 6.5 to 7.5 with 0.2 interval and the range of osmolality from 250 to 370 mOsm with 20 interval, and the period of sperm preincubation examined was 30, 60, 120, and 180 minutes. The ova developed to 2-cell embryos after 26hrs of incubation with preincubated sperm were evaluatated as in vitro fertilized ones. The results obtained were summarized as follows: 1. The optimal ranges of pH and osmolality of culture media and of sperm preincubation time for in vitro development of in vitro fertilized ova to blastocyst were pH 7.1 to 7.3, 250 to 350 mosmol and 60 to 180 min, respectively. 2. With the media of pH 7.1, 310 mOsm and sperm preincubation period of 120min in another experiment of large sample size, the in vitro fertilized ova was found 66.5% and the in vitro development of in vitro fertilized ova to blastocyst was found 35.8%. From the above results it was concluded that the optimal conditions of pH and osmolality of the media for mouse IVF and embryo culture, and the period of sperm preincubation might be 7.1, 310 mOam and 120min, respectively.

• Clinical and Experimental Reproductive Medicine
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• v.49 no.4
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• pp.225-238
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• 2022

The ultimate goal of human assisted reproductive technology is to achieve a healthy pregnancy and birth, ideally from the selection and transfer of a single competent embryo. Recently, techniques for efficiently evaluating the state and quality of preimplantation embryos using time-lapse imaging systems have been applied. Artificial intelligence programs based on deep learning technology and big data analysis of time-lapse monitoring system during in vitro culture of preimplantation embryos have also been rapidly developed. In addition, several molecular markers of the secretome have been successfully analyzed in spent embryo culture media, which could easily be obtained during in vitro embryo culture. It is also possible to analyze small amounts of cell-free nucleic acids, mitochondrial nucleic acids, miRNA, and long non-coding RNA derived from embryos using real-time polymerase chain reaction (PCR) or digital PCR, as well as next-generation sequencing. Various efforts are being made to use non-invasive evaluation of embryo quality (NiEEQ) to select the embryo with the best developmental competence. However, each NiEEQ method has some limitations that should be evaluated case by case. Therefore, an integrated analysis strategy fusing several NiEEQ methods should be urgently developed and confirmed by proper clinical trials.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.21 no.1
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• pp.1-18
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• 1995

We had tested the effect of amino propane sulfonic acid(APSA) on the human skin with non-invasive technique. It was tested with four parameters that were hydration, elasticity, color and skin wrinkles. In vitro culture systems, APSA stimulated the proliferation of fibroblasts bolt it didn't stimulate that of keratinocyte. Also we obtained the similar effects in the raft culture method. So we concluded that APSA affected the dermal region than the epidermal region. In clinical tests, APSA changed the skin color, pbiomechanical properties(especially elasticity) and reduced skin wrinkles of the volunteers. And we could get the better results of skin wrinkle improvement by use of Skin Visiometer than Silflo Image analysis systems.

• Korean Journal of Plant Resources
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• v.17 no.3
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• pp.257-264
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• 2004

Selection of stress-tolerant ginseng lines in fields is very difficult because it is almost impossible to control properly the environmental conditions of soil. On the contrary, it can be studied with ease to search for stress-tolerant ginseng lines through in vitro culture because of easy manipulation of stress conditions. This study was conducted for the selection of ginseng pure lines tolerant to salt stress. Murashige ＆amp; Skoog(MS) media with 2.5 folds of KNO$_3$, NH$_4$NO$_3$, MgSO$_4$.7$H_2O$, KH$_2$PO$_4$, and CaC1$_2$.2$H_2O$ was established for the selection of ginseng pure lines tolerant to salt stress in vitro. Among 88 ginseng pure lines bred by Korea Ginseng and Tobacco Research Institute, Punggi Hwangsuk, 78093, 82886, 78135, 86024 and KG104 lines was tolerant to salt stress. For the stable production of quality Korean ginseng, genetic tolerance to salt stress is one of important factors since relatively high salt concentrations in the ginseng nursery soil environment of Korea. Ginseng inbred pure lines were tested for their tolerance to salt stress through in vitro culture technique.

• Journal of Plant Biotechnology
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• v.7 no.1
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• pp.67-74
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• 2005

To overcome various disadvantages of conventional cul-ture vessels for micropropagation, a novel disposable vessel, the 'Vitron', made of a multi-layered $OTP^{(R)}$ film and supported by a polypropylene frame, was developed. The film possesses superior properties such as: high light transmittance, low water vapor transmittance and thermal stability and in particular, high gas-permeability. Single nodal explants, which were excised from the multiple shoots derived from shoot-tip culture, were cultured in Vitron and polycarbonate vessels on $3\%$ sugar-containing agar on MS medium and placed at 3000 ppm $CO_2$-enrichment at a low photosynthetic photon flux density (PPFD) ($45{\mu}mol\;m^{-2}\;s^{-1}$). The in vitro and ex vitro growth, and the net photosynthetic rate of in vitro and ex vitro plantlets were significantly enhanced in the Vitron compared to those cultured in a polycarbonate vessel. Explants that were cultured on the same MS medium under low PPFD at various $CO_2$ concentrations were also cultured at 3000 ppm $CO_2$- enrichment at various PPFD: 30, 45, 60, 75 and $90{\mu}mol\;m^{-2}\;s^{-1}$. The best in vitro and ex vitro growth obtained for 3000 ppm $CO_2$-enrichment at $75{\mu}mol\;m^{-2}\;s^{-1}$ PPFD. The novel Vitron vessel, when placed under the two conditions, may replace conventional culture vessels for the successful micropropagation of sweetpotato.

• Journal of Plant Biotechnology
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• v.30 no.3
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• pp.269-273
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• 2003

In order to develop an efficient micropropagation protocal for Eucalyptus pellita, on in vitro culture system has been was established by inducing axillary buds from greenhouse stock materials. Among 6different media tested, DKW medium was the best ot induce bast induce both shoot proliferation and growth. Average number of proliferated shoots of 403per explant was obtained at the concentration of 0.1mg/LBA. Most of the stem materials excreted phenolic compounds at the proximal part of the explant and caused darking of the media. Therefore, it was necessary to transfer frequently to a fresh medium and/or to add activated charcoal at the concentration of 0.02％(w/v). Generally on vitro roots were formed easily on 1/2DKW medium with NAA treatment. All the explants rooted at the medium containing 0.2mg/L NAA and displayed vigorous root growth in vitro culture conditions. After transferred to an artificial soil mixture (peatmoss: vermiculrite: perlite, 1:1:1, v/v/v) in the greenhouse, most rooted plantlets survived well without any morphological abnormalities. The results show that the species can be micropropagated effectively by the application of axillary bud culture system.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.2
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• pp.187-193
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• 2009

The present study identified the favorable conditions for isolation, enrichment and in vitro culture of highly purified, undifferentiated pig spermatogonial stem cell (SSC) lines that proliferate for long periods of time in culture. The colonies displayed morphology similar to miceSSC and were positive for markers of SSC (PGP9.5), proliferating germ cell (PigVASA), pre-meiotic germ cell (DAZL) and pluripotency (OCT4, SSEA-1, NANOG, and SOX2) based on immuno-cytochemistry and RT-PCR. The purity of these colonies was confirmed by negative expression of markers for sertoli cell (GATA4 and SOX9), peritubular myoid cell (${\alpha}$-SMA), differentiating spermatogonial and germ cells (c-KIT). The colonies could be maintained with undifferentiated morphology for more than two months and passaged more than 8 times with doubling time between 6-7 days. Taken together, we conclude that pigSSC could be successfully isolated and cultured in vitro and they possess characteristics similar to miceSSC.

• Journal of Embryo Transfer
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• v.17 no.3
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• pp.195-201
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• 2002

This study was to assess the developmental capacity of oocytes matured in vitro after 20, 15, 10, 5 and 0 days of organ culture when ovaries were isolated from juvenile mice at 0-, 5-, 10-,15- and 20-day old, respectively, and to develop in vitro culture system that observed a view to morphology of ovaries and nucleus maturation of oocytes. The size of ovaries decreased 35.9%, 8.7%, 1.2% and 14.4% after 20, 15, 10, 5 days of organ culture when the ovaries were isolated from 0-, 5-, 10 and 15-day old mice, respectively. After organ culture, the recovery rates, diameters of oocytes and the number of oocytes progressed from GV to MII were increased as increasing age of mice.

• The Plant Pathology Journal
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• v.36 no.6
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• pp.558-569
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• 2020

Fusarium head blight (FHB) is a devastating fungal disease of wheat (Triticum aestivum L.). The lack of genetic resources with stable FHB resistance combined with a reliable and rapid screening method to evaluate FHB resistance is a major limitation to the development of FHB resistant wheat germplasm. The present study utilized an immature wheat spike culture method to screen wheat spike culture derived variants (SCDV) for FHB resistance. Mycotoxin concentrations determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS) correlated significantly (P < 0.01) with FHB severity and disease progression during in vitro spike culture. Selected SCDV lines assessed for FHB resistance in a Fusarium field disease nursery in Carman, Manitoba, Canada in 2016 showed significant (P < 0.01) correlation of disease severity to the in vitro spike culture screening method. Selected resistant SCDV lines were also crossed with an elite cv. CDC Hughes and the progeny of F₂ and BC₁F₂ were screened by high resolution melt curve (HRM) analyses for the wheat UDP-glucosyl transferase gene (TaUGT-3B) single nucleotide polymorphism to identify resistant (T-allele) and susceptible (G-allele) markers. The progeny from the crosses were also screened for FHB severity using the immature spike culture method and identified resistant progeny grouped according to the HRM genotyping data. The results demonstrate a reliable approach using the immature spike culture to screen for FHB resistance in progeny of crosses in early stage of breeding programs.