• Clinical and Experimental Reproductive Medicine
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• 제28권2호
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• pp.121-129
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• 2001

Objective: The aim of this study is to compare the efficiency of a method for the cryopreservation of mouse blastocyst.. Methods: Mouse embryos were obtained at 2-cell stage and cultured to blastocyst stage in T6 medium supplemented with 10% fetal bovine serum. Morphologically normal blastocysts were collected and randomly divided to one control and four experimental groups. In control group, blastocysts were cultured in vitro continuously for additional two days. In group 2, blastocysts were exposed to vitrification solution (ethylene glycol) only without cryopreservation (exposure only group). In group 3, 4 and 5, blastocysts were cryopreserved by slow-freezing procedure with glycerol (slow-fteezing group) or by vitrification procedure using EM grids (EM grids group) and cryoloop (cryoloop group), respectively. Frozen blastocysts were thawed and cultured for additional two days. Twenty four hours after thawing, some blastocysts were fixed and stained with Hoechst 33342 (bisbenzimide) and the number of nuclei in each blastocysts were counted to confirm the survival of bias to cysts in experimental groups. Results: Survival rate and hatching rate of the blastocysts in slow-freezing group (24 h: 72.4% and 66.0%, 48 h: 63.2% and 64.6%) and EM grids group (24 h: survival rate 77.3%, 48 h: 70.1% and 71.4%) were significantly lower ($X^2$-test p<0.05) than those of control group (24 h: 93.4% and 86.0%, 48 h: 88.5% and 90.7%). In contrast, the survival rate and hatching rate of the blastocysts in cryoloop group (24 h: 84.1% and 84.1%,48 h 79.3% and 87.7%) is well compared with those in the control group. The mean (${\pm}SD$) cell number of blastocyst in the exposure only ($89.2{\pm}11.5$), EM grids ($85.0{\pm}10.3$) and cryoloop ($89.0{\pm}11.0$) groups, except slow-freezing group ($79.0{\pm}10.0$), were not significantly different from that of control group ($93.1{\pm}13.9$) 24 h after thawing (Student's t-test). Conclusion: This study demonstrates that higher survival rate of vitrified-thawed mouse blastocyst can be obtained using cryoloop as the embryo container at freezing rather than slow-freezing or vitrification using EM grids. The results of this study suggest that vitrification using cryoloop (with ethylene glycol) may be a preferable procedure for mouse blastocyst cryopreservation and could be applied to the human blastocyst cryopreservation.

• Clinical and Experimental Reproductive Medicine
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• 제24권3호
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• pp.281-292
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• 1997

The objective of this study was to compare retrospectively the survival and pregnancy rates(PR) of cryopresered-thawed embryos obtained from intracytoplasmic sperm injection (ICSI) or conventional in vitro fertilization (IVF). Ninety-six cycles of cryopresered-thawed embryo transfer (ET) were performed in 79 patients from June, 1996 to September, 1997 and grouped as followings: 20 cycles (16 patients) inseminated by ICSI (ICSI Group) and 76 cycles (63 patients) by conventional IVF (IVF Group). Slow-freezing and rapid-thawing protocol was used with 1.5M propanediol (PROH) and 0.1M sucrose as cryoprotectant. All embryos were frozen-thawed at the two pronuclear (2 PN) stage excluding four cycles in which the early cleavage stage embryos were frozen, and allowed to cleave in vitro for one day before ET. The duration from freezing to thawing was comparable in both groups ($mean{\pm}SD$, $112.1{\pm}80.0$ vs. $124.8{\pm}140.1$ days). The age of female ($31.2{\pm}3.4$ vs. $32.6{\pm}3.3$ years) and the endometrial thickness prior to progesterone injection ($9.4{\pm}2.0$ vs. $9.3{\pm}1.8$ mm) were also comparable in both groups. There was no significant difference in the outcomes of cryopreserved-thawed ET between two groups: survival rate ($85.2{\pm}16.1%$ vs. $82.2{\pm}19.7%$), cleavage rate ($96.9{\pm}6.7%$ vs. $94.7{\pm}13.0%$), cumulative embryo score (CES, $54.5{\pm}31.1$ vs. $49.0{\pm}20.0$), preclinical loss rate (5.0% vs. 5.3%), clinical miscarriage rate (0% vs 29.4%), clinical PR per transfer (35.0% vs. 22.4%), implantation rate (9.9% vs. 5.6%), and multifetal PR (42.9% vs. 17.6%). In conclusion, human embryos resulting from ICSI can be cryopreserved-thawed and transferred successfully, and the survival rate and PR are comparable to conventional IVF.

• 생물환경조절학회지
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• 제29권1호
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• pp.1-8
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• 2020

고구마 바이러스 무병묘의 기내급속증식을 위한 적정 생장 조절물질 및 sucrose 농도, 최소생장 기내보존(15℃)에 미치는 광의 영향과 생존율 및 기내생장 특성 등을 조사하였다. 고구마 무병묘의 마디배양은 0.2 mg·L^-1 BA 첨가배지에서 줄기신장, 줄기직경, 잎수, 뿌리수, 생체중 및 건물중 등이 가장 양호하였다. 배양부위 및 배지물리성에 따른 적정 sucrose 농도는 마디배양은 5% sucrose를 첨가한 고체배지에서, 정단배양은 3% sucrose를 첨가한 액체배지에서 줄기두께, 잎수, 뿌리수, 뿌리길이, 생체중 및 건물중 등의 생육에 가장 효과적이었다. 15℃ 저온항온기에서 고구마 무병묘의 최소생장 기내보존은 암상태에서는 3개월 내에 모두 고사하였으나, 적색:청색(7:3) 혼합 LED (150±5 μmol·m^-2·s^-1 PPFD)에서는 5개월까지 100% 생존하였다. 따라서 고구마 무병묘의 최소생장 기내보존에는 광이 필요하며, 샬레에 밀식(10 개체/샬레)할 경우, 좁은 공간에서 대량보존이 가능하였다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2002년도 춘계학술발표대회 발표논문초록집
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• pp.53-53
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• 2002

This study was carried out to verify the incidence of oocytes when vitrified at various maturation stages. Bovine cumulus-oocyte complexes were recovered from ovaries at a slaughter and then divided into five groups: control group(unvitrified oocytes), 0 hr. group(composed of oocytes vitrified before the onset of maturation) and 10, 14, and 20 hrs groups (vitrified respectively at 10, 14 and 20 hrs after the onset of maturation). (omitted)

• Asian Pacific Journal of Cancer Prevention
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• 제15권6호
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• pp.2405-2425
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• 2014

Radiation exposure leads to several pathophysiological conditions, including oxidative damage, inflammation and fibrosis, thereby affecting the survival of organisms. This review explores the radiation countermeasure properties of fourteen (14) plant extracts or plant-derived compounds against these cellular manifestations. It was aimed at evaluating the possible role of plants or its constituents in radiation countermeasure strategy. All the 14 plant extracts or compounds derived from it and considered in this review have shown some radioprotection in different in vivo, ex-vivo and or in vitro models of radiological injury. However, few have demonstrated advantages over the others. C. majus possessing antioxidant, anti-inflammatory and immunomodulatory effects appears to be promising in radioprotection. Its crude extracts as well as various alkaloids and flavonoids derived from it, have shown to enhance survival rate in irradiated mice. Similarly, curcumin with its antioxidant and the ability to ameliorate late effect of radiation exposure, combined with improvement in survival in experimental animal following irradiation, makes it another probable candidate against radiological injury. Furthermore, the extracts of P. hexandrum and P. kurroa in combine treatment regime, M. piperita, E. officinalis, A. sinensis, nutmeg, genistein and ginsan warrants further studies on their radioprotective potentials. However, one that has received a lot of attention is the dietary flaxseed. The scavenging ability against radiation-induced free radicals, prevention of radiation-induced lipid peroxidation, reduction in radiation cachexia, level of inflammatory cytokines and fibrosis, are some of the remarkable characteristics of flaxseed in animal models of radiation injury. While countering the harmful effects of radiation exposure, it has shown its ability to enhance survival rate in experimental animals. Further, flaxseed has been tested and found to be equally effective when administered before or after irradiation, and against low doses (${\leq}5Gy$) to the whole body or high doses (12-13.5 Gy) to the whole thorax. This is particularly relevant since apart from the possibility of using it in pre-conditioning regime in radiotherapy, it could also be used during nuclear plant leakage/accidents and radiological terrorism, which are not pre-determined scenarios. However, considering the infancy of the field of plant-based radioprotectors, all the above-mentioned plant extracts/plant-derived compounds deserves further stringent study in different models of radiation injury.

• Natural Product Sciences
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• 제9권2호
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• pp.60-63
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• 2003

Effect of grape seed extract (GSE) against Candida albicans was examined under in-vitro and in-vivo conditions. The GSE was extracted in ethanol. In-vitro results from an agar diffusion susceptibility assay showed the GSE inhibited C. albicans growth. This anticandidal effect was at dose-dependency. In experiments with animals, mice that received the GSE (0.5 mg per mice), intravenously (i.v.), before i.v.-infection wish viable C. albicans yeast cells survived longer than diluent (buffer)-received control mice. In contrast, when GSE was given to mice after the mice were infected with the yeast cells, these mice showed a similar survival rate as compared to control mice that received no treatment with the GSE. Taken together, these data indicate that GSE has prophylactic effect but not therapeutic effect against disseminated candidiasis.

• Plant Resources
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• 제7권1호
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• pp.1-6
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• 2004

In vitro proliferation system was achieved by using nodal segment excised from greenhouse grown juvenile stock plants of Alnus japonica. Stem explants were cultured on MS medium supplemented with different plant growth regulators of cytokinin and/or their combinations. The most effective cytokinin source was the combination of zeatin 2.0 mg/L and TDZ 0.05 mg/L producing the average number of shoots (16.8 $\pm$ 3.6). In addition, healthy roots were formed after small clumps of shoots were transferred to half strength of MS medium containing IBA 0.02 mg/L with optimal rooting capacity. Soil acclimatization was successfully conducted in cell tray containing artificially mixed soil with 92 % survival rate.

• 생물환경조절학회지
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• 제14권4호
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• pp.298-301
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• 2005

Potato (Solamum tuberosum 'Dejima') plantlets were investigated on culture type and initial quantity of inoculation in bioreactor and survival rate by hydroponics for mass production. rode stems (1 to 1.5cm in length) of potato plantlets multiplied in vitro were grown for 3 weeks in liquid Murashige and Skoog (MS) medium with sucrose $30 g\; L^{-1}$. When plantlets (80-node inoculation) were raised in 10L balloon type bubble (BB) bioreactor, the healthiest growth of plantlets was obtained from explants cultured in ebb & flow culture with medium supplied periodically 12 times per day. The suitable inoculation quantity of 20L BB bioreactor was 120 pieces of stem segments (mean 2.2g fresh weight) in ebb & flow culture. Number of nodal shoot was eight on the average. In controlled culture room, survival rate of plantlets at 7 days after stem cutting was above $70\%$ when they were acclimatized by hydroponics grown in deep flow and solid medium culture. The highest survival rate of the stem cutting plantlets was in nutrient solution adjusted to EC $1.4dS{\cdot}m^{-1}$. Stem cutting plantlets through one culture could be obtained $670\~900$, when plantlets were grown in ebb & flow culture during 3 weeks using a 20L bioreactor with initial 120 pieces of nodal segments. 11 is possible In do mass production of seedlings cultured in bioreactor and hydroponics.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2002년도 춘계학술발표대회 발표논문초록집
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• pp.41-41
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• 2002

Bovine cumulus-oocyte complexes were recovered from ovaries at a slaughter and then divided into five groups: control group(unvitrified oocytes), 0 hr. group(composed of oocytes vitrified before the onset of maturation) and 10, 14, and 20 hrs groups(vitrified respectively at 10, 14 and 20 hrs after the onset of maturation). The oocytes remained vitrified for 24 hrs, and then were thawed in 30℃ water bath. Survival and cleavage rates were defined as development rate on in vitro culture and stained with aceto-orcein or FDA test.

• 방사선산업학회지
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• 제4권4호
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• pp.307-312
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• 2010

Plants have evolved physiological, biochemical and metabolic mechanisms to increase their survival under the adverse conditions. This present study has been performed to select salt-tolerant rice mutant lines through in vivo and in vitro mutagenesis with gamma-rays. For the selection of the salt-tolerant rice mutants, we conducted three times of selection procedure using 1,500 gamma ray mutant lines resulted from an embryo culture of the original rice cv. Dongan (wild-type, WT): first, selection in the a nutrient solution with 171 mM NaCl; second, selection under in vitro condition with 171 mM NaCl; and third, selection in a reclaimed saline land. Based on a growth comparison of the entries, out of the mutant lines, two putative 2 salt tolerant (ST) rice mutant lines, ST-87 and ST-301, were finally selected. The survival rate of the WT, ST-87 and ST-301 were 36.6%, 60% and 66.3% after 7 days in 171 mM NaCl treatment, respectively. The WT and two salt tolerant mutant lines were used to analyze their genetic variations. A total of 21 EcoRI and Msel primer combinations were used to analyze the genetic relationship of among the two salt-tolerant lines and the WT using the ABI3130 capillary electrophoresis system. In the AFLP analysis, a total of 1469 bands were produced by the 21 primer combinations, and 700 (47.6%) of them were identified as having polymorphism. The genetic similarity coefficients were ranged from 0.52 between the ST-87 and WT to 0.24 between the ST-301 and the WT. These rice mutant lines will be used as a control plot for physiological analysis and genetic research on salt tolerance.