• Clinical and Experimental Reproductive Medicine
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• v.13 no.2
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• pp.121-127
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• 1986

We have reviewed 59 cases of patients amoung 65 cases who underwent IVF and ET with reasonable indications irom 1984 and the results as follows. 1. Major indications for IVF and ET were tubal factor (40.7%), unexplained infertility (25.4%), endometriosis (15.3%), failed AID and AIH (10.1 %), and sperm abnormality (8.5%). 2. For superovulation of human oocytes, l00mg of clomiphene citrate and 75 IU of HMG used. The monitoring of oocyte maturation was bone by ultrasound examination and serum 17-${\beta}$ estradiol, LH values. The peak $E_2$ value was 956.36${\pm}$702.13 pg/ml. 3. The oocytes were obtained by laparoscopy 24-36 hours after the injection of HCG. 4. The mean numbers of follicles at laparoscopy was 3.06 and the successful rate of laparoscopy was 79.7%. 5. And 165 follicles were aspirated from which 98 oocytes were recovered, 59.4% of all follicles had at least one oocyte aspirated. 21.4% of the eggs were mature, 52.0% were moderate, 26.5%. were immature. 6. 67.3% of oocytes were cleaved and were transferred at 4-6 cell stages. 7. Four pregnancies including one chemical pregnancy and one spontaneous abortion were established by ${\beta}$-subunit, u-hCG and ultrasound examinations.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2000.10b
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• pp.16-17
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• 2000

Aralia elata is found in mountain areas all over Korean peninsula. Aralia elata is the scientific name for Japanese angelica tree. The tree belongs to the family Araliaceae, commonly known as ginseng family. Bud sprouts from apical shoot tip of the plants are rich in flavor and thus mainly used for both folk medicine and vegetable. The stalks with apical buds are gathered in the early spring and planted in sandy soil or water in the greenhouse. The sprouting buds are then collected and sold as fresh vegetable. Although the plants have been used for food, they have been cultivated in a very small scale. In spring, local farmers just go around mountain areas to search the trees and gather the stalks as much as they get and sell them to the market. No conservation efforts have been made to stop the exploitation or to save the dwindling population. We tried to provide local farmers with the plants that may be used as an alternative to stalks from wild populations. This will hel! p conserve the wild populations. However, it is hard to propagate them either by conventional cuttings or by seed germination in a short period of time. Mass propagation using tissue culture systems have shown a great promise with several woody plants. Recently we developed a mass propagation technique via somatic embryogenesis system using mature and/ or juvenile explants for Aralia elata. Several factors affecting somatic embryogenesis system including SE(somatic embryo) induction, embryogenic callus proliferation, SE germination, plant regeneration and transplanting to field will be presented. And some problems arising for the somatic embryogenesis system will be also discussed.lso discussed.

• Clinical and Experimental Reproductive Medicine
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• v.40 no.4
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• pp.148-154
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• 2013

Objective: To compare the mouse oocyte vitrification outcomes of the CryoLogic vitrification method (CVM) and the conventional open method using a Cryotop. Two CVM methods (original CVM and modified CVM) were tested. Methods: Mature oocytes obtained from female BDF-1 mice were vitrified by two-step exposure to equilibrium and vitrification solutions. Three vitrification protocols were tested on three groups: the CVM-kit, modified CVM, and Cryotop groups. After exposure to the two solutions, the oocytes were vitrified. After warming, the oocytes were fertilized in vitro, and the embryo development was assessed. Blastomeres positive for caspase were counted using an in situ assay kit. The spindle morphology and chromosome configurations of warmed vitrified oocytes were also assessed. Results: The modified CVM and Cryotop groups showed similar developmental capacities, and similar proportions of cells with intact spindles and chromosome configurations. The modified CVM protocol was superior to the original CVM protocol for developmental competence and intact spindle preservation. However, the CVM group showed a relatively higher number of apoptotic cells in blastocysts. Conclusion: Closed vitrification using the modified CVM protocol may be used as an alternative to the conventional open method, but strategies to decrease apoptosis in the blastomere need to be investigated.

• Molecules and Cells
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• v.40 no.8
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• pp.558-566
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• 2017

Regular monitoring on experimental animal management found the fluctuation of ART outcome, which showed a necessity to explore whether superovulation treatment is responsible for such unexpected outcome. This study was subsequently conducted to examine whether superovulation treatment can preserve ultrastructural integrity and developmental competence of oocytes following oocyte activation and embryo culture. A randomized study using mouse model was designed and in vitro development (experiment 1), ultrastructural morphology (experiment 2) and functional integrity of the oocytes (experiment 3) retrieved after PMSG/hCG injection (superovulation group) or not (natural ovulation; control group) were evaluated. In experiment 1, more oocytes were retrieved following superovulation than following natural ovulation, but natural ovulation yielded higher (p < 0.0563) maturation rate than superovulation. The capacity of mature oocytes to form pronucleus and to develop into blastocysts in vitro was similar. In experiment 2, a notable (p < 0.0186) increase in mitochondrial deformity, characterized by the formation of vacuolated mitochondria, was detected in the superovulation group. Multivesicular body formation was also increased, whereas early endosome formation was significantly decreased. No obvious changes in other microorganelles, however, were detected, which included the formation and distribution of mitochondria, cortical granules, microvilli, and smooth and rough endoplasmic reticulum. In experiment 3, significant decreases in mitochondrial activity, ATP production and dextran uptake were detected in the superovulation group. In conclusion, superovulation treatment may change both maturational status and functional and ultrastuctural integrity of oocytes. Superovulation effect on preimplantation development can be discussed.

• Journal of Crop Science and Biotechnology
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• v.10 no.2
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• pp.92-97
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• 2007

Antibiotics used for suppressing Agrobacterium in plant transformation procedure might have negligible effects on plant tissues and regeneration. The effects of antibiotics on growth suppression of Agrobacterium and plant regeneration were investigated for enhancing Agrobacterium-mediated transformation using wheat mature embryos. Antibiotics tested, except carbenicillin, were able to suppress that embryos were coated with a layer of Agrobacterium cells in callus induction medium. Agrobacterium growth was suppressed minimally at 50 mg/l of timentin, while cefotaxime and clavamox were completely suppressed at relative high concentration of 250 mg/l. In the treatment of carbenicillin, initiation of growth suppression of Agrobacterium occurred at 750 mg/l of concentration because Agrobacterium KYRT1 contains the carbenicillin resistant gene. In Agrobacterium inoculation, effects of antibiotics were significantly different on the rate of callus induction and shoot formation. Almost embryos were induced calli at 50 mg/l of timentin whereas callus induction rate was achieved above 90% at 100 mg/l and 250 mg/l of cefotaxime and clavamox, respectively. Shoot formation rate was higher in the treatment of timentin than that of cefotaxime and clavamox at 500 mg/l of concentration, respectively. Timentin can be used as a good antibiotics in Agrobacterium-mediated wheat transformation.

• Development and Reproduction
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• v.21 no.4
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• pp.457-466
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• 2017

Survival of embryos largely depends on yolk processing during early development. Proteolytic enzymes, cathepsin B & D (ctsb & ctsd) are known to have some important roles in yolk processing of various fish species. Mature female red spotted groupers were injected with human chorionic gonadotropin (HCG) to induce ovulation. The fertilized eggs and embryos were sampled at 0, 4 and 24 HPF (hours post fertilization). Survivals of each groups of embryos were checked at 24 and 48 HPH (hours post hatching). Transcripts of ctsb & ctsd showed the highest level at 0 HPF and relatively high at 4 HPF, but greatly decreased at 24 HPF. In bad egg quality group (BE, embryos survived until 24 HPH), transcript level of ctsb at 4 HPF were significantly lower than the transcript level at the same stage in good egg quality group (GE, embryos survived until 48 HPH) while no significant change of ctsb transcript level was observed at 0 or 24 HPF between BE and GE. Transcript level of ctsd was decreased at 24 HPF, but the difference was not as strong as the case of ctsb transcript. These results suggest that maternal ctsb transcript rather than ctsd transcript is likely to be involved in egg quality resulting in the difference of survival rate of embryos at early developmental period in this species.

• Forest Science and Technology
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• v.14 no.4
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• pp.160-168
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• 2018

The seeds of Elaeocarpus serratus, a tropical underutilized fruit tree are characterized by hard seed coat and consequent poor water uptake and low germination. To improve the regeneration through seeds, various parameters such as viability of seeds, water uptake, and effect of seed mass on germination and pretreatments were performed using a completely randomized design (CRD). Tetrazolium (TZ) test was conducted using fresh, mature seeds revealed $50{\pm}2.56%$ mean viability. Seeds of different weight classes showed similar pattern of water uptake and the saturation level was achieved at 60 hrs of soaking. Seeds belong to weight class 2.6-3.5g were germinated ($12.5{\pm}1.26%$) with $175{\pm}1.75days$ (d) of mean time taken for germination (MTG). Germination capacity of seeds varied significantly among different populations and Varkala population gave $12.5{\pm}1.1%$ germination with $174.6{\pm}2.5d$ MTG. Among various seed treatments, mechanical scarification was superior in germination and significant reduction in MTG ($p{\leq}0.05$). The mechanical scarification by complete removal of seed coat resulted in $49.2{\pm}1.52%$ germination within a short period of time ($9.52{\pm}0.89d$ MTG). However, the complete removal of seed coat without damaging to embryo is a difficult task. An alternate treatment (Mechanical scarification II) by making cracks on nut faces vertically followed by soaking in distilled water for 24 hrs gave $48.4{\pm}1.73%$ germination with significantly reduced MTG ($12.14{\pm}0.56d$) over unsoaked, untreated control ($6.5{\pm}1.84%$ germination and $197.18{\pm}1.79d$ MTG; $p{\leq}0.05$). This treatment (Mechanical scarification II) is therefore recommended for E. serratus seeds as it can adopt easily and can achieve 7 fold increases in germination over control. The recorded germination through mechanical scarification is in tune with realized viability percentage of the seeds.

• Korean Journal of Ichthyology
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• v.34 no.1
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• pp.16-24
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• 2022

The eggs development and early life history of Sarcocheilichthys variegatus wakiyae was investigated. The adult fishes were collected in Yeongsangang River, Damyang-gun, Jeollanam-do, Korea, and their spawning inducement was carried by sex maturation hormone (Ovaprim) injections. Spawning amount was mean 31±3.5 Mature eggs were cicular in shape and mean 2.40±0.05 mm in diameter. Hatching period of the embryo was 230 to 240 hours after fertilization under water temperature of 22℃. The newly hatched larvae were 10.7±0.19 mm in total length (TL). 3 days after hatching, postlarvae was measured 12.0±0.13 mm in TL and the tip of the caudal fin was bent at 45°. 11 days after hatching, it was measured 14.6±0.13 mm in TL, and transferred to juvenile as the number of fins (iii7 dorsal fin, iii6 anal fin, i7 ventral fins) reached a constant number of each part.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.32 no.3
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• pp.277-286
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• 1987

Abscisic acid (ABA) content of the seed and endocarp during stratification were analyzed and then examined in relation to the embryo growth and germination. In mature red fruitlet, the ABA content was remarkably higher in sarcocarp than those in both seed ans endocarp. During the stratification before dehiscence, ABA content was gradually decreased in both seed and endocarp. After 90 days(dehiscent percentage; 96%) it came to 90 pmol/ g DW(69% decrease) and to 41 pmol/ g DW (80% decrease) in seed and in endocarp, respectively. The ratio of free from to total ABA content showed constant decrease in seed, but remained at higher level in endocap than in seed. Correlation between the decrease of ABA content and embryo growth showed higher significance in seed than in endocarp. During the stratification after dehiscence, ABA content in seed was gradually decreased at 4$^{\circ}C$ and 15$^{\circ}C$, After 90 days it came to 28 pmol/ g DW (69% decrease) and to 46 pmol/ g DW (49% decrease) at 4$^{\circ}C$ and at 15$^{\circ}C$, respectively. The ratio of free form to total ABA content was gradually increased at 4$^{\circ}C$, but remained almost constant at 15$^{\circ}C$. Correlation between the decrease of ABA content and days to first germination showed positive singificance only at 4$^{\circ}C$, whereas the correlation between the decrease and mean germination percentage per day showed negative significance at 4$^{\circ}C$, but positive significance at 15$^{\circ}C$. The above results indicate the ABA of the seed end endocarp during the stratification before dehiscence seems to be concerned with the immature embryo growth, but that of the seed during the stratification after dehiscence seems to show little effect on the germination capability(degree of breaking physiological dormancy).

• Clinical and Experimental Reproductive Medicine
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• v.26 no.2
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• pp.171-178
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• 1999

The follicular fluid (FF) of ovary contains various biological active products which affected on the growth of follicles and the fertilization of oocyte in physiological reproductive process of mammals. This study was designed to determine the effects of human FF on fertilization of oocyte and embryonal development in vitro culture. The FF was prepared as clear without blood contamination by needle aspiration from mature follicles of human at the time of oocytes retrieval for in vitro fertilization (IVF). As the medium for culture in vitro of embryonal cells, human tubal fluid (HTF) supplemented with follicular fluids at concentrations of 10%, 40% and pure FF were used. These effects were compared to control group of cultured embryos in HTF supplemented with 0.4% BSA (bovine serum albumin). For IVF, 64 eggs in control group, 67 eggs in 10% FF, 57 eggs in 40% FF and 64 eggs in pure FF were respectively allocated. And the rates of fertilization were almost similar in all groups as resulting 82.81% in control, 85.07% in 10% FF, 87.71% in 40% FF and 81.25% in pure FF. On the examination for embryonal cleavage from fertilized eggs, the rates of developing to 4 cell stage was similar in all groups, as results 98.11% in control, 98.27% in 10% FF and 98% in 40% FF but 78.84% in pure FF. And the rates of developing to 8-16 cell stage were significantly reduced as 44% in 40% FF and 44.23% in pure FF (p<0.05) compare to 71.69% in control media. As likewise, the rates of developing to morular stage were also significantly reduced to 36% (p<0.05) and 21.15% (p<0.01) respectively in 40% FF and pure FF. And the rates to blastocystic stage of embryo was lowest as 7.69% in pure FF (Table 1). The quality of embryonal cells on cleavage to the 8-16 cell stage was poorer, higher concentrations of FF. The rates of grade 1 in pure FF, as 23.07%, was lowest compare to those of other groups, in which the rates of grade 1 in control, 10% FF and 40% FF were 58.49%, 47.36% and 34% respectively. And on the contrary, the rate of grade 4 in pure FF was highest as 23.07%, while those were 5.66% in control, 8.77% in 10% FF and 20% in 40% FF (Table 2). On the viability of embryos, the rate of embryonal cell death was more rise, at the higher concentrations as well as longer exposure in the follicular fluid. At 48 hours after in vitro culture of embryos, the rate of survival embryos in pure FF was markedly lowered as 44.23%, compare to that of control (p<0.05). But there was not significant difference between the rates of survival embryos in each group beside the pure FF, which the rates were 77.35% in control, 70.17% in 10% FF and 60% in 40% FF respectively. And at 72 hours after in vitro culture, the rates of survival embryos were also significantly dropped to 21.15% in pure and 36% in 40% at concentration of FF compare to 62.26% in control (p<0.05, p<0.01). Finally, the rate of embryonal death at 96 hours after in vitro culture was highest as 82.69% in pure FF among all groups which those were 35.84 in control, 56.14% in 10% FF and 64% in 40% FF respectively (Fig. 1, 2, 3). In conclusion, this study suggests that the FF has no effects, in particular, to the in vitro fertilization of oocytes but exerted a bad effect to the cleavage, quality and viability of the embryonal cells during in vitro culture. However, the FF is harmful on embryonal development at conditions in higher concentration and especially on the embryos after $8{\sim}16$ cell stage.