• Reproductive and Developmental Biology
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• v.30 no.4
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• pp.301-305
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• 2006

The aim of this study was to investigate whether addition of porcine epididymal fluid (pEF) into culture medium during in vitro maturation influences the nuclear maturation of porcine germinal vesicle (GV) oocytes. Porcine cumulus-oocyte complexes (COCs) from follicles were cultured in tissue culture medium 199 (TCM 199) containing pEF. After 48hr of culture, oocytes were examined for evidence of GV breakdown, metaphase I, anaphase-telophase I, and metaphase II. The proportion of oocytes reaching at metaphase II (M II) stage was significantly (p<0.05) increased in oocytes cultured in the media supplemented with 10% pEF during in vitro maturation than in those without pEF regardless of cumulus presence or absence (54.6% vs 22.5%, 51.7% vs 24.2%). The supplementation of pEF during maturation of oocyte enhanced oocytes maturation in a dose-dependent manner in vitro. Also significant differences (p<0.05) in the percentage of MII oocytes were observed according to exposure period in pEF. Present study suggests that pEF contains a enhancing component(s) for nuclear maturation of porcine immature oocytes in vitro.

• Development and Reproduction
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• v.25 no.3
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• pp.173-183
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• 2021

The incidence of infertility among individuals of reproductive age has been growing due to genetic and environmental factors, and considerable research efforts are focused on solving this issue. Ovarian development is an overly complex process in the body, involving the interaction between primordial germ cells and gonad somatic cells. However, follicles located in the center of the in vitro ovary are poorly formed owing to ovarian complexity, nutrient deficiency, and signaling deficiency. In the present study, we optimized methods for dissociating gonads and culture conditions for the in vitro generation of miniaturized ovaries. The gonads from embryos were dissociated into cell masses and cultured on a Transwell-COL membrane for 3-5 weeks. Approximately 12 follicles were present per in vitro ovary. We observed that miniaturized ovaries successfully matured to MII oocytes in vitro from 150 to 100 ㎛ gonad masses. This method will be useful for investigating follicle development and oocyte production.

• Korean Journal of Animal Reproduction
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• v.14 no.2
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• pp.115-124
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• 1990

These experiments were undertaken as a basic study to develop immunocontraceptive vaccine and to understand the role of zona pellucidae in early fertilization process by investigating the effect of monoclonal and polyclonal antibody to porcine zona pellucidae and polyclonal antibody to mouse zona pellucidae on the fertilization of porcine and mouse eggs in vitro. The results obtained in these experiments were summarized as follows : 1. Treatment of porcine and mouse eggs with undiluted anti-zona serum produced intense precipitation layer on the poricne and mouse zonae, respectively, thus resulting in the total inhibition of sperm adherence on surface of zona. 2. In vitro fertilization of eggs pre-treated with 0.3∼10% of various antibodies was examined, and resulting in that 5 and 10% of rabbit polyclonal antibodies to porcine zona inhibited completely both in vitro fertilization and polyspermy of porcine eggs while monoclonal to porcine zona and rabbit polyclonal antibody to mouse zona did not inhibit in vitro fertilization but monoclonal antibody reduced the rate of polyspermy compared to that of control group. Almost the same results were obtained in the study on the effect of anti-zona serum on in vitro fertilization of mouse eggs.

• Journal of Plant Biotechnology
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• v.49 no.3
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• pp.230-239
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• 2022

Holostemma annulare (Family Asclepiadaceae) is an invaluable vulnerable medicinal plant; the root tubers are used in Ayurveda medicine and by folk healers to treat various ailments. In this study, Schenk and Hildebrandt medium fortified with the cytokinins 6-benzyl adenine, kinetin, and auxins, including indole 3-butyric acid, indole 3-acetic acid, α-naphthaleneacetic acid, and 2,4-dichlorophenoxyacetic acid, were checked for their efficiency on root tuber induction from different explants. Adventitious root tubers were more successfully induced from in vitro leaf segments and shoots when cultured in Schenk and Hildebrandt medium supplemented with 0.5 mg/l of α-naphthaleneacetic acid. In addition, preliminary phytochemical analysis of in vitro root tubers and identification of different secondary metabolites were conducted. Thin layer chromatography and high performance thin layer chromatography analysis of the crude methanolic extracts of the in vitro root tuber identified the presence of lupeol, a bioactive triterpene. Adventitious root tuber induction offers a novel method for the in vitro production of bioactive metabolites that can be scaled up by bioreactors, thus ensuring the conservation and sustainable utilization of H. annulare. The study warrants further scale-up production and pharmacological investigation that can be extended for pharmaceutical needs.

• Animal Bioscience
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• v.34 no.1
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• pp.56-65
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• 2021

Objective: This study was conducted to investigate the effects of molasses and Lactobacillus plantarum on the ensiling quality and in vitro rumen fermentation of sudangrass silage prepared with or without wilting. Methods: The ensiling experiment, measured with 3 replicates, was carried out according to a 2×4 (wilted stages×additives) factorial treatment structure. Dry matter of the fresh (210 g/kg fresh matter) or wilted (305 g/kg fresh matter) sudangrass were ensiled (packed into 5.0-L plastic jars) without additive (control) or with molasses (M), Lactobacillus plantarum (LP), or molasses + Lactobacillus plantarum (M+LP). After 60 days of ensiling, the silages were analyzed for the chemical, fermentation, and in vitro characteristics. Results: After 60 days of ensiling, the fermentation parameters were affected by wilted, the additives and the interactions of wilted with the additives (p<0.05). The M+LP treatment at wilted had higher lactic acid levels and V-score (p<0.05) but lower pH values and butyric acid concentrations than the other treatments. In comparison with sudangrass before ensiling, after ensiling had lower dry matter and higher non-fibrous carbohydrate. The in vitro gas production, in vitro dry matter digestibility, in vitro crude protein digestibility, and in vitro acid fiber detergent digestibility changed under the effects of the additives. Significant interactions were observed between wilted and the additives in terms of in vitro gas production at 48 h, asymptotic gas production, gas production rate, half time, and the average gas production rate. The total volatile fatty acid levels in the additive treatments were higher than those in the control. Conclusion: Wilting and supplementation with molasses and Lactobacillus plantarum had the ability to improve the ensiling quality and in vitro nutrient digestibility of sudangrass silage. The M+LP treatment at wilted exhibited the strongest positive effects on silage quality and in vitro ruminal fermentation characteristics.

• Journal of Embryo Transfer
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• v.11 no.2
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• pp.137-144
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• 1996

The present study was carried out to obtain the pregnancy and delivery rate following transfer of fresh and frozen-thawed Korean native cattle(KNC) blastocysts(1~4 em-bryos / head) produced in vitro to Holstein recipients. The pregnancy rate of fresh and frozen-thawed KNC blastocysts produced in vitro was 50%(7 /14 heads) and 38.5%(5 /13 heads), respectively. The pregnancy rate of frozen-thawed KNC blastocysts produced in vitro frozen using 1.5M ethylene glycol and 1.4M glycerol for cryoprotectant was 33.3%(2 /6 heads) and 42.9 %(3 /7 heads), respectively. Seven calves including 2 sets of twin were born fiom 5 pregnant recipients receiving eleven fresh blastocysts. Three pregnant recipients were aborted among four pregnant recipients receiving twelve frozen-thawed blastocysts and one calf was born from the rest of one pregnant recipient.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.28 no.3
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• pp.171-184
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• 2002

The present study was undertaken to develop the in vitro sunprotection factor(SPF) measurement method having good correlation with in vivo method. As in vitro method, 8% homomentyl salicylate, P3 reference standard and sunscreens on market were measured using SPF 290 analyzer and were compared the SPF with labed value. In vitro SPF of 8% HMS and P3 reference standard were 4.59 $\pm$ 0.12 and 14.94 $\pm$ 0.83. There are good correlation, correlation factor were 0.9506 and 0.9769 respectively, between the in vitro and in vivo SPF for the sunscreen creams and lotions examined. Correlation factors of makeup base & liquid goundation, lotion labled with “shake before use”, compact powder were 0.8812, 0.8632 and 0.5984. The best sample applied method of compact powder was 1:0.8 mixture with cream base. These results suggest that the in vitro test method could be used as an alternative method for SPF.

• Korean Journal of Veterinary Research
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• v.36 no.4
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• pp.919-927
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• 1996

In the last few years, methods for in vitro culture of early embryo stages from oocytes matured and fertilized in vitro using suitable cell culture systems have been established. But the factors affecting pregnancy rates following transfer of bovine embryos produced in vitro were not evaluated enough. So this study was performed to investigate the effects of quality and stage of embryos, parity and Corpus Luteum quality of recipients on pregnancy rates following non-surgical transfer of bovine embryos produced in vitro. Oocytes aspirated from small antral follicles of ovaries obtained at a local slaughter house were matured, fertilized with frozen-thawed semen and co-cultured for 6-7 days by utilizing co-culture system with bovine oviduct epithelial cell in vitro. After co-culture, embryos were transfered to recipients on day 7 (estrus=day 0). Recipients were monitored by ultrasonic scanning method or observation for estrus and rectal palpation after 50 days from transfer. The results of this study are follows. 1. Of the 70 recipients, 70%(49 of 70) had not showed estrus sign between day 0 and day 50, but 22.9%(16 of 70) was diagnosed not pregnant. Therefore the overall pregnancy rate of this study was 47.1%(33 of 70). 2. The pregnancy rate of recipients transfered with excellent(66.7%) and good(54.5%) embryos were higher than that of recipients transfered with fair embryos(15.8%) (p<0.05). 3. The pregnancy rate of recipients transfered with morula, compacted morula, blastocyst and expanded blastocysts were 46.2, 55.0, 62.5 and 50.0%, respectively. 4. The pregnancy rates of recipients transfered to heifer and cow were 54.5 and 55.2%, respectively. 5. The pregnancy rates of recipients with CL score I, II(66.7, 63.6%) were higher than those of recipients with CL score III (10%), (p<0.05). Success of transfer of embryos produced in vitro depends on many variables. The important factors identified in this study were the quality of embryos and the CL score of recipient animals after non-surgical transfer of embryos matured, fertilized and cultured in vitro.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.19 no.1
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• pp.57-76
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• 1993

Phototoxicity is a complex phenomenon which may involve photochemical reaction and biological response mechanism. This complexicity and iii mal protecting tendency has led to the development of various in-vitro approaches as sensitive, alternative test to the in-vivo phototoxicity test. In this study, we investigated not only the sensitivity of two microorganism, (C. albicans and 5. typhimurium TA 98 about UV) but also a correlation between in-vitro and in-vivo phototoxicity test using UV A and 1 Furthermore, we studied the effect of irradiation method which were as follows 1) irradiate to material and microorganism, simultaneously 2) irradiate to only material 3) irradiate to material and microorganism, respectively In each irradiation method, it showed no significant difference, However we were able to observe the more sensitive phototoxicity in S. typhimurium TA 98 than C. albicans, and the results of in-vitro test using 5. typhimurium TA 98 had a good correlation with those of in-vivo test.

• Journal of Pharmaceutical Investigation
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• v.38 no.3
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• pp.183-189
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• 2008

The present study compared the feasibility of Caco-2 and MDCK cells as an efficient in-vitro model for the drug classification based on Biopharmaceutics Classification System (BCS) as well as an in-vitro model for drug interactions mediated by P-gp inhibition or P-gp induction. Thirteen model drugs were selected to cover BCS Class I{\sim}IV$ and their membrane permeability values were evaluated in both Caco-2 and MDCK cells. P-gp inhibition studies were conducted by using vinblastine and verapamil in MDCK cells. P-gp induction studies were also performed in MDCK cells using rifampin and the P-gp expression level was determined by western blot analysis. Compared to Caco-2 cells, MDCK cells required shorter period of time to culture cells before running the transport study. Both Caco-2 and MDCK cells exhibited the same rank order relationship between in-vitro permeability values and human permeability values of all tested model compounds, implying that those in-vitro models may be useful in the prediction of human permeability (rank order) of new chemical entities at the early drug discovery stage. However, in the case of BCS drug classification, Caco-2 cells appeared to be more suitable than MDCK cells. P-gp induction by rifampin was negligible in MDCK-cells while MDCK cells appeared to be feasible for P-gp inhibition studies. Taken all together, the present study suggests that Caco-2 cells might be more applicable to the BCS drug classification than MDCK-cells, although MDCK cells may provide some advantage in terms of capacity and speed in early ADME screening process.