• Reproductive and Developmental Biology
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• 제34권1호
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• pp.21-25
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• 2010

The purpose of this study was to investigate the effects of the collection time, co-culture and sperm penetration of canine oocytes on in vitro maturation and fertilization. The oocytes were cultured in TCM-199 media containing hormonal supplements (10% FCS, 10 IU/ml HCG, 10 IU/ml PMSG) at 5% $CO_2$, 95% air, $38^{\circ}C$. The in vitro maturation rate to MII stage of in vitro oocytes recovered from ovaries that collected at follicular, luteal and inactive phases of the reproductive phase for 44~72 hrs were 19.2%, 12.2%, and 6.0%, respectively. Follicular phases oocytes had a significantly higher in vitro maturation rate than oocytes collected at luteal and anestrus stage (p<0.05). The in vitro maturation rates to the MII stage of canine oocytes after 48 hrs of culture with glutathione, pyruvate, or glutathione + pyruvate were 12.5%, 10.7%, and 17.5%, respectively. This was higher than that in both alone or the combination of the two compared to the control group (19.0%). The sperm penetration rates of in vitro matured oocytes by fresh and frozen semen were 29/80 (36.3%) and 18/80 (22.5%), respectively. Although there are limited reports about canine oocytes co-culture and in vitro fertilization, our results on in vitro maturation is comparable to the results from other researches.

• 한국가축번식학회지
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• 제21권3호
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• pp.275-280
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• 1997

This study was carried out to investigate on the survival and in vitro developmental rates of bisected bovine embryos by microblade, micropipette and pronase methods. Bisected embryos cultured for 1∼7 days in TCM-199 media with 10 FCS＋hormones. Survival and in vitro developmental rates was defined on in vitro culture or FDA-test. The results are summarized as follows ; 1. The survival and in vitro developmental rates of bisected bovine embryos by microblade, micropipette and pronase methods were 22.2, 16.7, 15.0% and 22.2, 23.3, 18.8%, respectively. In vitro developmental rate of bisected bovine embryos was significantly lower than that of non-bisection embryos(27.8% and 25.0%). 2. In vitro developmental rates of bovine embryos bisected for 1, 2, 4, 8, 16 cells stages during in vitro culture in 10% FCS＋TCM-199 media were 25.0, 20.0, 20.0, 15.0 and 6.7%, respectively. 3. In vitro developmental rates of intact and free-zona pellucida of bisected demi-embryos during in vitro culture in 10% FCS＋TCM-199 media were 25.6, 16.7%, respectively. 4. In vitro developmental rates of biopsied embryos and biopsied blastomeres during in vitro culture in 10% FCS＋TCM-199 media were 20.0, 11.1%, respectively.

• 한국가축번식학회지
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• 제22권1호
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• pp.101-104
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• 1998

This study examined the effects of growth factors in TCM199 on bovine 1-cell embryos development in vitro. After 6 day to 11 day in culture, 15.8%(19/120), 15.3%(20/130), 21.8%(35/160), 27.0%(56/207), 26.3%(53/201) and 30.7%(40/130) of the 1-cell embryos developed into expanding blastocysts in su, pp.ementing TCM199 with control, insulin, IGF-I, IGF-II, FGF and EGF, respectively. Hatching rate of 1-cell embryos in su, pp.ementing TCM199 with FGF, EGF and IGF-II were 21.4%(53/247), 20.3%(42/206) and 16.8%(41/243), respectively. The beneficial effect of growth factors on embryo development in vitro could be duplicated. These data indicate that the presence of FGF, EGF or IGF-II in the culture medium is beneficial for embryo development in vitro and accelerate cell differentiation.

• Journal of Pharmaceutical Investigation
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• 제32권4호
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• pp.321-325
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• 2002

IVIVC (In vitro/in vivo correlation) is useful for predicting in vivo results from in vitro data. The aim of this study was to develop IVIVC of sustained release diltiazem. For this purpose, three types of diltiazem tablets with different in vitro dissolution rates were prepared. An in vitro dissolution testing method comprising of paddle apparatus, 50 rpm, water as dissolution medium was developed. Under these condition, we demonstrated that AUCinf could be predicted by evaluating $d_{70%}$ (time dissolved 70%) in vitro since the in vivo AUCinf was correlated with the in vitro $d_{70%}$ (r=-0.9981).

• Asian-Australasian Journal of Animal Sciences
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• 제21권1호
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• pp.59-65
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• 2008

Six total mixed rations (TMR) containing 0, 4, 6, 8, 10, 12% tannin (TMR I-VI), using Accacia nilotica pods as a source of tannin, were used to study the effect of Acacia tannin on in vitro nutrient digestibility and gas production in goats. This study also investigated the degraded products of Acacia nilotica tannin in goat rumen liquor. Degraded products of tannins were identified using high performance liquid chromatography (HPLC) at different hours of incubation. In vitro digestibility of dry matter (IVDMD) and organic matter (IVOMD) were similar in TMR II, and I, but declined (p<0.05) thereafter to a stable pattern until the concentration of tannin was raised to 10%. In vitro crude protein digestibility (IVCPD) decreased (p<0.05) with increased levels of tannins in the total mixed rations. Crude protein digestibility was much more affected than digestibility of dry matter and organic matter. In vitro gas production (IVGP) was also reduced (p<0.05) with increased levels of tannins in the TMR during the first 24 h of incubation and tended to increase (p>0.05) during 24-48 h of incubation. Gallic acid, phloroglucinol, resorcinol and catechin were identified at different hours of incubation. Phloroglucinol and catechin were the major end products of tannin degradation while gallate and resorcinol were produced in traces. It is inferred that in vitro nutrient digestibility was reduced by metabolites of Acacia nilotica tannins and ruminal microbes of goat were capable of withstanding up to 4% tannin of Acacia nilotica pods in the TMR without affecting in vitro nutrient digestibility.

• 한국축산식품학회지
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• 제33권6호
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• pp.689-695
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• 2013

This study was conducted to investigate the effects of in vitro human digestion on the antioxidant activity of blueberry leaf extracts (BLE) in emulsion-type sausages (ETS). Leaves from four cultivars of blueberries (Bluecrop, Bluegold, Duke, and Northland) collected from a wild blueberry farm were extracted with 80% ethanol. ETS were prepared with 0.2% BLE. The samples were then passed through an in vitro human digestion system which simulates the composition of the mouth, stomach, and small intestine juice. Only one phenolic compound (chlorogenic acid) was detected in the BLE. Northland BLE had appreciably higher amounts of chlorogenic acid than that of other BLE, both before and after in vitro human digestion. Antioxidant activity of any BLE was not influenced by in vitro human digestion, whereas the antioxidant activity of chlorogenic acid standard increased in response to in vitro human digestion in both 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and ferric-reducing ability of plasma (FRAP). In the present study, the antioxidant activities of the BLE were not strongly influenced by in vitro human digestion, and the antioxidant activity depended on the chlorogenic acid content of ETS. Thus, compounds from blueberry leaves may have important applications in the future as natural antioxidants for meat products.

• 한국동물생명공학회지
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• 제34권1호
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• pp.35-39
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• 2019

This study aimed to recover the ovarian function through in vitro culture of preantral follicles from aged mice. First, we isolated the preantral follicles from ovaries of sixty-seven-week old B6D2F1 mice with decreased fecundity to know how many follicles were present in them, which was 6 preantral follicles including 2 primary, 2 early secondary and late secondary follicles from 8 aged mice. It was confirmed that a few follicles (~2) were present in aged mice through histological analysis compared to adult mice as control. The 9 days of in vitro culture of preantal follicles showed in vitro growth and induced maturation after treatment with hCG (2.5 IU/mL) and EGF (5 ng/mL). Cumulus cells in the cumulus-oocyte complexes (COCs) were removed using hyaluronidase and oocytes at the germinal vesicle (GV) and GV breakdown (GVBD) were obtained from preantral follicle culture of aged mice in vitro. In conclusion, these observations demonstrated that there still were a few preantral follicles in the ovaries of 67 week-old mice, which we were able to culture in vitro and oocytes were obtained from them. This study proposed an in vitro culture system using preantral follicle as a therapeutic strategy for fertility preservation in humans for assisted reproductive medicine.

• 약학회지
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• 제44권5호
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• pp.424-431
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• 2000

A biopharmaceutics drug classification system for correlation between in vitro dissolution and in vivo bioavailability is proposed based on recognizing that drug dissolution and gastrointestinal permeability are the fundamental parameters controlling the rate and extent of drug absorption. The objective of this study was to assess whether in vitro dissolution profiles of immediate-release beta-blocker tablets can be correlated with intestinal membrane permeability and/or in vivo bioavailability In vitro dissolution of the beta-blocker tablets was examined using KP VII Apparatus II methods at various pH. Intestinal membrane permeability was determined in vitro using the diffusion chamber method. Bioavailablity parameters were cited from literatures. The dissolution profiles did not accurately represent the in vivo bioavailablity However there were good correlations between intestinal membrane permeability and log P (noctanol/buffer). The correlations obtained in this study indicated that in vitro diffusion chamber method could be used to predict intestinal absorption in vivo.

• Asian-Australasian Journal of Animal Sciences
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• 제24권11호
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• pp.1541-1546
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• 2011

This study was carried out to examine the influence of minimum essential medium (MEM) vitamins supplementation to in vitro maturation medium and in vitro culture medium on the development of porcine embryos. Porcine embryo development was investigated following cultivation in both in vitro maturation and culture medium with the supplementation of MEM vitamins (0, 0.1, 0.2 and 0.4%) using immature oocytes collected from the ovary of prepubertal gilts. Embryo development was observed and the total cell number in each blastocyst generated under the culture conditions was quantified following supplementation of the medium. The embryonic development rate of the blastocyst and hatched blastocyst was higher, but not significantly so, when 0.4% MEM vitamins were supplemented to the in vitro maturation medium of the porcine oocyte. Interestingly, the total number of cells in the blastocyst was significantly higher in the in vitro maturation MEM vitamins supplemented group compared to either the untreated group or the group which had MEM vitamins supplemented to both in vitro maturation and in vitro culture medium simultaneously (p<0.05). Therefore, the supplementation of 0.4% MEM vitamins to the in vitro mature medium has a beneficial effect on the embryonic development of in vitro produced blastocysts derived from the immature porcine oocytes.

• Asian-Australasian Journal of Animal Sciences
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• 제14권11호
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• pp.1645-1646
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• 2001

The objective of this study was to develop an in vitro digestion technique to estimate bioavailable energy values of different corn hybrids in swine and poultry. A total of 21 samples were investigated; 18 normal corn (NC) and 3 high-oil corn (HOC) hybrids. One step-two enzymes digestion system was introduced to develop the in vitro technique. The gross energy (GE) values of NC hybrids were correlated with the in vitro disappearing energy values (IVE; r=0.85, p<0.01), the in vitro energy digestibilities (IVED; r=0.70, p<0.01), and the in vitro DM digestibilities (IVDM; r=0.55, p<0.05). It appears, however, that IVE values of NC and HOC hybrids were not significantly different according to the one step-two enzyme digestion system. Results of in vivo and in vitro estimates suggested that there was no significant correlation between them in poultry. The IVE value was regressed linearly with ME and DE values in swine with low regression coefficient (34 and 41%, respectively).