• 한국수정란이식학회지
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• 제15권1호
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• pp.39-46
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• 2000

This study was conducted to investigate the effect of hormones, protein sources and anti-oxidants on in vitro maturation (IVM) and in vitro fertilization(IVF) of bovine follicular oocytes. The rates of Holstein follicular oocytes classified as grade A and B(50.2% and 33.2%) were higher than those of Hanwoo cattle(40.3% and 32.0%, P<0.05). The cumulus cell expansion rates of oocytes cultured in TCM-199 and Ham's F-10 medium supplemented with 10% FCS and hormones were higher (81.9~87.6%) than those of non-treated groups (74.5~81.7%). The fertilization rates of oocytes cultured in TCM-199 and Ham's F-10 medim supplemented with 10% FCS, 1% BSA and 10% bFF was 53.8~55.0%, 51.4~52.6%, and 47.0~50.0%, respectively. The polyspermy rates was 13.6~14.2%, 10.0~11.1%, and 10.0%, respectively. When the oocytes were cultured in TCM-199 and Ham's F-10 medium with 50${\mu}{\textrm}{m}$ $\alpha$-tocopherol, the fertilization rates was 62.0 and 60.2%, respectively. In the maturation medium added of 100${\mu}{\textrm}{m}$ cysteamine, the fertilization rates was 64.7 and 66.7%, respectively. The fertilization and polyspermy rates of treated groups were higher than those of non-treated group. The results show that hormones, protein sources and anti-oxidants can provide a benefit for in vitro maturation and fertilization of bovine follicular oocytes.

• 한국수정란이식학회지
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• 제23권2호
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• pp.77-80
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• 2008

In the present study, effects of concentration of cryoprotectant solutions on the nuclear maturation of vitrified-thawed porcine oocytes were examined. Also, the developmental capacity of vitrified-thawed immature porcine oocytes following ICSI was investigated. Oocytes were cultured in NCSU-23 medium supplemented with 5% FBS at $38^{\circ}C$ in 5% $CO_2$ and air. The in vitro maturation rate of vitrified-thawed oocytes ($24.1{\pm}2.5%$) was lower than that of the control ($46.0{\pm}3.2%$, p<0.05). The in vitro maturation rate of vitrified-thawed oocytes treated with $1.0{\sim}5.0\;ug$ CB + NCSU- 23 medium were $22.2{\pm}3.0%$, $30.7{\pm}3.2$, $46.3{\pm}3.1%$, $38.5{\pm}3.2%$, respectively. The in vitro maturation rate ($46.3{\pm}3.4%$) of the vitrified-thawed oocytes treated with $3.0\;{\mu}g$ CB for 30 min was the highest of all vitrification groups. When the in vitro developmental rates of the vitrified-thawed (with EDS and EDT) oocytes following ICSI were $18.5{\pm}2.5%$, $16.4{\pm}2.1%$, respectively. This results were lower than the control group ($24.0{\pm}2.5%$).

• 한국수정란이식학회지
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• 제23권2호
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• pp.93-100
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• 2008

This study was conducted to establish an in vitro maturation (IVM) system by selection of efficient macromolecule in the porcine in vitro production (IVP) technology. To choose the efficient macromolecules in the development of porcine embryos, the effects of 3 kinds of macromolecules (porcine serum; PS, porcine follicular fluid; pFF, and polyvinyl alcohol; PVA) supplemented in IVM media on the maturation, cleavage, and development rates to blastocyst of parthenogenetic activation (PA) and in vitro fertilization (IVF) embryos were examined. The maturation rates of porcine oocytes in media supplemented with PS were significantly higher than those with pFF and PVA (92.4% vs. 85.4%, 77.1%; p<0.05). In the cleavage and development to blastocyst rates, supplement with PS or pFF in the IVM media was more effective than PA. However, there were no significant differences in cleavage and development to blastocyst between PS and pFF group. From the results of this study, it was demonstrated that PS was optimal macromolecule in the porcine IVM media.

• Asian-Australasian Journal of Animal Sciences
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• 제29권3호
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• pp.352-358
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• 2016

Quercetin (QT) and taxifolin (TF) are structurally similar plant-derived flavonoids that have antioxidant properties and act as free radical scavengers. The objective of this study was to investigate effects of QT and TF on nuclear maturation of porcine oocytes. Effects of TF at 0, 1, 10, and $50{\mu}g/mL$ on oocyte nuclear maturation (polar body extrusion) were investigated. After incubation for 44 h, there were no significant differences between the treatment and control groups except in the $50{\mu}g/mL$ group which was significantly lower (59.2%, p<0.05) than the other groups (control: >80%). After parthenogenetic activation, further in vitro development of QT- or TF-treated vs control oocytes was investigated. A significantly higher proportion of QT-treated ($1{\mu}g/mL$) oocytes developed into blastocysts compared to controls (24.3% vs 16.8%, respectively); however, cleavage rate and blastocyst cell number were not affected. The TF-treated group was not significantly different from controls. Levels of reactive oxygen species (ROS) and intracellular glutathione (GSH) in oocytes and embryos in a culture medium supplemented with QT or TF were measured. Both treatment groups had significantly lower (p<0.05) levels of ROS than controls, however GSH levels were different only in QT-treated oocytes. We conclude that exogenous flavonoids such as QT and TF reduce ROS levels in oocytes. Although at high concentration ($50{\mu}g/mL$) both QT and TF appear to be toxic to oocytes.

• 대한치과보철학회지
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• 제33권2호
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• pp.317-334
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• 1995

The purpose of this study was to compare the bone formation, maturation around HA-and titaniumcoated dental implants in dog. 5 hydroxyapatite coated IMZ implants and 5 titanium coated IMZ implants were placed into the previously extracted site in the mandible of 5 adults dogs. All dogs were injected intravenously Tetracycline, Alizalin red S, and Calcein for bone fluorescent labelling, After the experimental period of 16weeks, the dogs were sacrificed and tissue samples around the implants were obtained. Microscopic observations(ligth, polariged and fluorescence microscope), morphometric analysis, line profile with EPMA, and quantitative analysis for Ca,P, and Ti were performed. The results were as follows ; 1. Bone maturations around the implants were relatively lower than those of natural teeth. No significant differences in bone maturation and remodeling patterns were observed between the two implants groups. 2. Calcification of bone surrounding the implants was initiated in 8-11 weeks for HA-coated implants, while it took 11 weeks or more for Ti-coated implants. 3. Bone-to-implants contact ratio of 82.63% was recorded for HA-coated group and 72.25% for titanium coated group, with no significant difference between the two groups. 4. Bone around the implants exhibited reduced quantity of Ca and P in the $100{\mu}m$ region relative to natural teeth, while the rest of the regions showed no statistical differences. No significant differences were found between the two implant groups. 5. There was a separation of HA layer from the implant core and subsequent infiltration of inflammatory cells into the resulting space in the HA-coated implants, and evidences of phagocytosis of HA particles by macrophages. Bone calcification was more rapid around HA-coated implants compared to titanium-coated implants, but HA coated implants did not show any significant differences either in the degree of calcification or the bone-to-implant contact ratio over Ti coated implants. HA coated implants may have complications associated with HA absorption and separation of HA layer from the implant core.

• 한국가축번식학회지
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• 제19권3호
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• pp.181-189
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• 1995

In this study, methods on fabrication of microtool and setting of micromanipulator were examined and relationship between first polar body extrusion rate and maturation time of follicular oocyte, enulceation rae and repetition of trial, and enucleation rate and maturation period were investigated. The results are as follows: 1. Suitable outside diameter of micropipette tube was 1mm. Holding pipette with less than diameter of oocyte was fitred for manipulation, and zona dissection needle was easily operated when its sharp-point had diameter of about 8 ${\mu}{\textrm}{m}$ and length of 300${\mu}{\textrm}{m}$. The injection pipette with 20~35${\mu}{\textrm}{m}$ outside diameter was adequate for injection of blastomere into perivitelline space. 2. Separation of blastomere was effective when zona pellucida had cut with zonadissection needle and the embryo was pipetted gently with the pipette that had narrower diameter than that of embryo until separation of blastomeres had completed. 3. The extrusion rate of first polar body was 78% during 20~24% hours incubation for maturation. 4. According to repetitions of micromanipulation, the enucleation rate was increased to 85% and the time required for enucleation of a oocyte was shortened to 3 min. 5. The extrusion rate of first polar body and enucleation rate were 82 and 76% respectively, in the group of the oocytes cultured for 22 hours. However in the group cultured for 24 hours, the extrusion rate of first polar body and enucleation rate were 53 and 100% respectively.

• Asian-Australasian Journal of Animal Sciences
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• 제26권4호
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• pp.501-508
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• 2013

The effects of amino acids and dipeptides on in vitro production of porcine embryos and accumulation of ammonia in culture medium during developmental stages were examined in this study. The maturation, fertilization and development of embryonic cultures were performed in modified Tissue culture medium (mTCM)-199 supplemented with 10% (v/v) porcine follicular fluid, modified Tyrode's albumin lactate pyruvate (mTALP) medium, and modified North Carolina State University (mNCSU)-23 medium, respectively. In addition, amino acids and dipeptides of different concentrations and combinations were used to treat the embryos. The addition of L-alanyl-L-glutamine (AlnGln)+L-glycyl-L-glutamine (GlyGln) significantly (p<0.05) improved oocyte maturation, fertilization and the incorporation and oxidation of 14C(U)-glucose when compared to the control group and other treatment groups. Additionally, 2-4 cell, 8-16 cell, morula and blastocyst development increased significantly (p<0.05) following treatment with AlnGln+GlyGln when compared to the control group and other treatment groups, while this treatment reduced the accumulation of ammonia. Taken together, these findings suggest that treatment with AlnGln+GlyGln may play an important role in increasing the rate of porcine oocyte maturation, fertilization and embryonic development by reducing the level of accumulated ammonia measured in the culture media.

• Clinical and Experimental Reproductive Medicine
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• 제14권1호
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• pp.7-17
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• 1987

It appears that a major determinant of the success of in vitro fertilization is the selection of the optimal follicle containing an oocyte capable of being fertilized and producing a normal pregnancy. However, the hormonal basis of oocyte maturation is not well substantiated by the as yet available informations. It has been suggested that prolactin(PRL) may stimulate the formation of an oocyte maturation inhibitor and thus inhibit the maturation of oocyte. During the hyperstimulated menstrual cycles serum estradiol($E_2$) levels are markedly elevated, and it seems justified to assume that serum prolactin levels may be elevated since estrogens are potent stimulators of prolactin secretion. This study was carried out to ascertain the effect of the elevated serum estradiol levels on the serum prolactin levels in women undergoing ovarian hyperstimulation with either hMG and/or clomiphene citrate. Serum estradiol and prolactin profiles were measured from third menatrual cycle day to ovulation or ovum aspiration day in 11 normal menstruating women and 30 women who underwent an in vitro fertilization procedure with ovarian hyperstimulation by hMG, clomiphene citrate/hMG, clomiphene citrate. Ovum aspiration was performed 36 hours after hCG administration. The day of ovum aspiration or ovulation was designated Day 0. Serum estradiol levels increased progressively during the follicular phase and this rise peaked on Day-1 at a mean concentration of 1,204${\pm}$189.0pg/ml in Group II(hMG), 1,194${\pm}$167.9pg/ml in Group III(clomiphene citrate/hMG), 1,035${\pm}$195.1pg/ml in Group IV(clomiphene citrate) respectively and on Day -2 of 336${\pm}$34.5pg/ml in Group I(normal control). The elevated estradiol levels fen rapidly after ovulation or ovum aspiration. Serum estradiol values of hyperstimulated groups(Group II, III, IV) were significantly higher than that of control group(Group I) from Day -6 to Day +1, but there was no significant difference of estradiol values among the hyperstimulated groups. Serum prolactin levels increased and peaked on Day +1 at a mean concentration of 60.8${\pm}$14.4ng/ml in Group II, 34.2${\pm}$7.0ng/ml in Group III, 30.1${\pm}$5.7ng/ml in Group IV respectively, but no significant elevation was observed in Group I. Levels of estradiol and prolactin can be positively and significantly correlated in the hyperstimulated groups. However, the increase of serum prolactin levels in hMG group was significantly higher than those in clomiphene citrate/hMG or clomiphene citrate group.

• 한국수정란이식학회지
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• 제32권4호
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• pp.257-265
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• 2017

Ferulic Acid (FA) is a metabolite of phenylalanine and tyrosine, a phenolic compound commonly found in fruits and vegetables. Several studies have shown that FA has various functions such as antioxidant effect, prevention of cell damage from irradiation, protection from cell damage caused by oxygen deficiency, anti-inflammatory action, anti-aging action, liver protective effect and anti-cancer action. In this study, we investigated the maturation rate, intracellular glutathione (GSH) and reactive oxygen species (ROS) of porcine oocytes by adding FA to the in vitro maturation (IVM) medium and examined subsequent embryonic developmental competence at 5% oxygen through parthenogenesis. There is no significant difference between the control group ($0{\mu}M$) and treatment groups ($5{\mu}M$, $10{\mu}M$, $20{\mu}M$) on maturation rates. Intracellular GSH levels in oocyte treated with $5{\mu}M$ of FA significantly increased (P < 0.05), and $20{\mu}M$ of FA revealed significant decrease (P < 0.05) in intracellular ROS levels compared with the control group. Oocytes treated with FA exhibited significantly higher cleavage rates (79.01% vs 89.19%, 92.20%, 90.89%, respectively) than the control group. Oocytes treated with $10{\mu}M$ showed significantly higher blastocyst formation rates (28.3% vs 40.3%, respectively) after PA than the control group. Total cell numbers in blastocyst of $10{\mu}M$ FA displayed significantly higher (39.4 vs 51.9, respectively) than the control group. In conclusion, these results suggested that treatment with FA during IVM improved the developmental potential of porcine embryos by increasing intracellular GSH synthesis and reducing ROS levels. Also, there was an improvement of cleavage rate, blastocyst formation and total cell numbers in blastocysts. It might be associated with Keap1-Nrf2 pathway as an antioxidant regulate pathway that plays a crucial role in determining the sensitivity of cells to oxidative damages by regulating the basal and inducible expression of enzymes which is related to detoxification and anti-oxidative effects, stress response enzymes and/or proteins and ABC transporters.

• 한국패류학회지
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• 제26권2호
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• pp.151-156
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• 2010

본 연구는 말백합 산업화를 위하여 어미성숙 관리한 인위성숙개체군과 시기별로 어미를 수송 채란한 자연성숙개체군을 산란유발 방법 별로 산란을 유도하여, 수정율, 부화율, D형 유생 발생률을 조사하였고, 유생사육 방법별로 유생의 성장과 생존율 등을 조사하였다. 인위성숙개체군의 간출시간별로 산란 유도한 결과 간출 4시간 및 8시간이 반응률이 23% 및 32%로 비교적 높았고, 수온 상승별로는 $28^{\circ}C$ 이상 되어야 반응이 시작되었다. 간출 및 수온상승 병행 시험구에서는 간출 4시간 및 $28^{\circ}C$에서 반응률이 45% 이상으로 나타났고, 29, 30, $31^{\circ}C$에서는 유의한 차이를 보이지 않아 말백합 어미는 간출시간보다는 수온상승에 반응률이 높은 것으로 나타났다. 어미수송개체군의 시기별 산란유발 결과는 2009년 8월 6일이 반응률 67.6%, D형 유생 발생률이 96%로 가장 높게 나타났다. 유생사육 시험 중 수온별로는 유생의 성장과 생존율을 고려할 때 유생사육 적정수온은 $27-31^{\circ}C$, 최적수온은 $29^{\circ}C$로 나타났고, 염분은 25에서 성장 및 생존율이 비교적 높았으며, 수용밀도별로는 5 inds./mL에서 성장 및 생존율이 비교적 높게 나타났음을 알 수 있었다.