• Proceedings of the KSAR Conference
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• 2000.10a
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• pp.9-9
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• 2000

The mammalian ovary has a large number of primordial and preantral follicles, which are a potential source of oocytes for the in vitro mass production of embryos. Several in vitro culture systems have been developed to support the growth and development of oocytes from mouse preantral follicles. Under the appropriate condition, meiotically incompetent oocytes from preantral follicles can grow to final size and complete nuclear maturation in vitro. Furthermore, the successful production of live young from in vitro grown and matured oocytes demonstrates that oocytes from preantral follicles are able to acquire full developmental capacity in vitro. However, the efficiency of in vitro production of embryos from mouse preantral follicles is still low. In farm animals as well as human, the growth of oocyte from preantral follicle to the meiotic competence stage has yet to be demonstrate. Therefore, further studies to improve the culture condition or to develope new culture system should be needed in the future. In addition, the visible progress in the establishment of the in vitro culture system for preantral follicles of farm animals and human could help to enlarge the populations of valuable agricultural, phamaceutical product-producing, and endangered animals, and to rescue the oocytes of women about to undergo clinical procedures that jeopardize oocytes.

• The Korean Journal of Physiology and Pharmacology
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• v.23 no.4
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• pp.231-236
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• 2019

In drug discovery or preclinical stages of development, potency parameters such as $IC_{50}$, $K_i$, or $K_d$ in vitro have been routinely used to predict the parameters of efficacious exposure (AUC, $C_{min}$, etc.) in humans. However, to our knowledge, the fundamental assumption that the potency in vitro is correlated with the efficacious concentration in vivo in humans has not been investigated extensively. Thus, the present review examined this assumption by comparing a wide range of published pharmacokinetic (PK) and potency data. If the drug potency in vitro and its in vivo effectiveness in humans are well correlated, the steady-state average unbound concentrations in humans [$C_{u_-ss.avg}=f_u{\cdot}F{\cdot}Dose/(CL{\cdot}{\tau})=f_u{\cdot}AUCss/{\tau}$] after treatment with approved dosage regimens should be higher than, or at least comparable to, the potency parameters assessed in vitro. We reviewed the ratios of $C_{u_-ss.avg}$/potency in vitro for a total of 54 drug entities (13 major therapeutic classes) using the dosage, PK, and in vitro potency reported in the published literature. For 54 drugs, the $C_{u_-ss.avg}$/in vitro potency ratios were < 1 for 38 (69%) and < 0.1 for 22 (34%) drugs. When the ratios were plotted against $f_u$ (unbound fraction), "ratio < 1" was predominant for drugs with high protein binding (90% of drugs with $f_u{\leq}5%$; i.e., 28 of 31 drugs). Thus, predicting the in vivo efficacious unbound concentrations in humans using only in vitro potency data and $f_u$ should be avoided, especially for molecules with high protein binding.

• Journal of Embryo Transfer
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• v.9 no.3
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• pp.235-241
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• 1994

The study was conducted to determine the optimal hormone and glucose levels during the in vitro culture of bovine oocytes matured and fertilized in vitro for blastocyst development. Oocytes matured in TCM 199 + 10% FCS + hormones and glucose were fertilized in vitro in a TALP medium with swim up separated and heparin-treated epididymal cauda spermatozoa. Oocytes were cultured for 2~5 days in synthetic oviduct fluid medium (SOFM) supplemented with 10% FGS and with different hormone and glucose levels, and further cultured 5 days same medium in SOFM. The results are summarized as follows : The in vitro maturation and penetration rates of porcine oocytes cultured in TCM 199 media containing PMSG, hCG, PMSG + hCG, hCG + $\beta$ estradiol, PMSG + $\beta$ estradiol 0 to20 hours after insemination were 88.0% and 81.8%, 82.6% and 68.4%, 80.0% and 75.0%, 80.0% and 65.0%, 77.3% and 64.7%, respectively. The in vitro maturation and penetration rates of porcine oocytes cultured in TCM 199 media containing PMSG, hCG, PMSG + hCG, hCG + $\beta$ estradiol, PMSG + $\beta$ estradiol 20 to 40 hours after insemination were 92.0% and 87.0%, 92.0% and 82.6%, 91.3% and 81.0%, 85.2% and 73.9%, 87.5% and 81.0%, respectively. The cleavage and in vitro developmental rates to blastocyst of porcine oocytes cultured in TCM 199 media containing 0.05 mM, 0.10 mM, 0.30 mM, 0.50 mM, 1.00 mM, and 3.00 mM glucose lelvels 0~3 days after insemination were 31.5~48.1% and 10.0~16.7%, respectively. The cleavage and in vitro developmental rates to blastocyst of porcine oocytes cultured in TCM 199 media containing 0.05 mM, 0.10 mM, 0.30 mM, 0.50 mM, 1.00 mM, and 3.00 mM glucose levels 4~8 days after insemination were 30.0~53.8% and 8.7~19.2%, respectively. The cleavage and in vitro developmental rates to blastocyst were higher in TCM 199 media containing various glucose levels 0~3 days after insemination than 4~8 days.

• Animal Bioscience
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• v.35 no.10
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• pp.1592-1605
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• 2022

Objective: The objective of this study was to select an effective in vitro digestion-fermentation model to estimate the effect of decreasing dietary crude protein (CP) on odor emission during pig production and to suggest potential prediction markers through in vitro and in vivo experiments. Methods: In the in vitro experiment, three diet formulations with different CP contents (170 g/kg, 150 g/kg, and 130 g/kg) but containing the same standardized ileal digestible essential amino acids (SID-EAA) were assessed. Each diet was evaluated by two different in vitro gastric-intestinal phase digestion methods (flask and dialysis), combined with fresh pig feces-ferment inoculation. Eighteen growing barrows (31.9±1.6 kg) were divided into three groups: control diet (180 g CP/kg, without SID-EAA adjustment), 170 g CP/kg diet, and 150 g CP/kg diet for 4 weeks. Results: The in vitro digestion results indicated that in vitro digestibility was affected by the gastric-intestinal phase digestion method and dietary CP level. According to the gas kinetic and digestibility results, the dialysis method showed greater distinguishability for dietary CP level adjustment. Nitrogen-related odor compounds (NH₃-N, indole, p-cresol, and skatole) were highly correlated with urease and protease activity. The feeding study indicated that both EAA-adjusted diets resulted in a lower odor emission especially in p-cresol and skatole. Both protease and urease activity in feces were also closely related to odor emissions from nitrogen metabolism compounds. Conclusion: Dialysis digestion in the gastric-intestinal phase followed by fresh fecal inoculation fermentation is suitable for in vitro diet evaluation. The enzyme activity in the fermentation and the fecal samples might provide a simple and effective estimation tool for nitrogen-related odor emission prediction in both in vitro and in vivo experiments.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.1
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• pp.33-38
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• 2004

This study was aimed at testing the gene expression of antioxidant enzymes and apoptosis genes for in vitro culture in porcine embryos produced by in vitro maturation/in vitro fertilization (IVM/IVF). Pocine preimplantation embryos obtainted from IVM/IVF can be successfully culture in vitro, but they are delayed or stop to develop at specific developmental stage. Many factors such as reactive oxygen species and apoptosis in an IVM/IVF system followed by in vitro culture influence the rate of production of viable blastocysts. Porcine embryos derived from IVM/IVF were cultured in the atmosphere of 5% $CO_2$ and 20% $O_2$ at $38.5^{\circ}C$ in NCSU23 medium. The patterns of gene expression for antioxidant enzymes and apoptosis genes during in vitro culture in pocine IVM/IVF embryos were examined by the modified semi-quantitative single cell reverse transcriptase-polymerase chain reaction (RT-PCR). Porcine embryos produced by in vitro procedures were expressed mRNAs for CuZn-SOD, GAPDH and GPX, whereas transcripts for Mn-SOD and catalase were not detected at any developmental stages. Expression of caspase-3 mRNA was detected at 2 cell, 8 cell 16 cell and blastocyst, but p53 mRNA was not detected at any stages. The fas transcripts was only detected in blastocyst stage. These results suggest that various antioxidant enzymes and apoptosis genes play crucial roles in vitro culture of porcine IVM/IVF embryos.

• Journal of Forest and Environmental Science
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• v.25 no.2
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• pp.119-126
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• 2009

An efficient in vitro plant regeneration system was developed through shoot proliferation from axillary buds of Tectona grandis (L.f), APNBV-1 (Andhra Pradesh North Badrachalam Venkatapuram-1) clone. Multiple shoots of high quality were produced in vitro from axillary bud explants. An average of 4.39 shoots/explant were obtained on Murashige and Skoog's (MS) medium supplemented with plant growth regulators (PGRs) benzyl amino purine (BA), kinetin (KN), indole acetic acid (IAA), gibberillic acid ($GA_3$), growth adjuvants casein hydrolysate (CH), adenine sulphate (Ads) and antioxidants ascorbic acid, polyvinyl pyrrollidine (PVP). Eighty five percent of rooting was observed in ex vitro rooting media containing IBA and vermiculite. In ex vitro rooting, single shoots with 2 to 3 nodes were subjected to IBA of different concentrations at different periods of time intervals. Direct rooting in vermiculite at 500 ppm concentration of IBA resulted in 4.3 number of roots with 2 cm length. Minimum response of rooting and length of roots were recorded at 100 ppm concentration of IBA. Planlets were transferred to plastic bags for short acclimatization stage in green house where they survived at 95%.

• Korean Journal of Animal Reproduction
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• v.14 no.4
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• pp.253-261
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• 1990

This study was carried out to investigate the factors affecting development in vitro of follicular oocytes fertilized in vitro in Korean Native Cattle. The bovine ovaries were obtained at a slaughter house and the follicular oocytes were recovered by aspirating the follicular fluid from the visible follciles of 3~6mm. The bovine oocytes were matured in vitro for 20~24 hours in TCM0-199 containing 10% FCS and hormones (0.02AU/ml FSH, 10$\mu\textrm{g}$/ml LH, 1$\mu\textrm{g}$/ml estradiol-17$\beta$). The matured oocytes were fertilized in vitro using Percoll-separated frozen-thawed spermatozoa in BO solution containing caffeine(5mM) and heparin(10$\mu\textrm{g}$/ml). Twenty-four hours after insemination, the oocytes were cultured in vitro and then the effects of cumulus cell layer, co-culture with cumulus cells, bovine oviduct epithelial cells from ampulla or isthmus on development of ova, were studied. The results obtained are summarized as follows : 1. The in vitro development degree of oocytes attached with compact and dense layered cumulus cells was higher than that with 3~4 layered cumulus cells to be 9~16cells(P<0.01). 2. When the in vitro fertilized oocytes were co-cultured with bovine oviduct epithelial cells or cumulus cells, the development rate to be morula was 20.2% and 12.7%, respectively and the rates were higher than that of control, 2.1%(P<0.05). 3. The development rate to be morula was 15.8% and 23.8%, respectively when the in vitro fertilized oocytes were co-cultured with bovine oviduct epithelial cells from ampulla or isthmus, and the rates were higher than that of control, 0%(P<0.05%).

• The Journal of Korean Academy of Prosthodontics
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• v.56 no.4
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• pp.391-395
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• 2018

Increasing demands for zirconia material in clinics, assessment of biocompatibility of zirconia is essential. In this article, a review of in vitro studies of zirconia compatibility was performed. Zirconia showed great biocompatibility at in vitro studies with various cell lines such as fibroblasts, osteoblasts, and lymphocytes. Many studies reported that zirconia caused no cytotoxicity or mutation. Zirconia also showed less bacterial adhesion. There were no adverse effects except for small reduced strength with in vitro study mimicking long-term exposure of body fluid. According to the study with ostoblast-like cells, zirconia could regulate genes of immunity, molecular transport, and cell cycle. Such gene regulating was considered as one of the reasons of zirconia biocompatibility. With biocompatibility of zirconia powders, in vitro studies had controversial conclusions. It seems that zirconia powders might have cytotoxicity.

• Korean Journal of Animal Reproduction
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• v.16 no.3
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• pp.209-215
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• 1992

These studies were carried out to overcome 2-cell block and in vitro development to blastocysts in vitro fertilization of mouse embryos. The unfertilized ova were obtained by superovulation in ICR mice of 4 to 6 weeks old. Tyrode's 280 solution was used as basal media, and the pH range of media examined was designed from 6.5 to 7.5 with 0.2 interval and the range of osmolality from 250 to 370 mOsm with 20 interval, and the period of sperm preincubation examined was 30, 60, 120, and 180 minutes. The ova developed to 2-cell embryos after 26hrs of incubation with preincubated sperm were evaluatated as in vitro fertilized ones. The results obtained were summarized as follows: 1. The optimal ranges of pH and osmolality of culture media and of sperm preincubation time for in vitro development of in vitro fertilized ova to blastocyst were pH 7.1 to 7.3, 250 to 350 mosmol and 60 to 180 min, respectively. 2. With the media of pH 7.1, 310 mOsm and sperm preincubation period of 120min in another experiment of large sample size, the in vitro fertilized ova was found 66.5% and the in vitro development of in vitro fertilized ova to blastocyst was found 35.8%. From the above results it was concluded that the optimal conditions of pH and osmolality of the media for mouse IVF and embryo culture, and the period of sperm preincubation might be 7.1, 310 mOam and 120min, respectively.

• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.133-142
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• 2002

The objective of this study was to examine maturation, fertilization and developmental rate of the in vitro-grown mouse oocytes, and to compare these results with those of oocytes grown and matured in vivo. The preantral follicles isolated from 12-day-old mice were cultured on Transwell-COL membrane inserts. After in vitro growth and maturation, 72.5 % of oocytes grown in vitro produced polar body which can be comparable to in vivo growth (70.5 %). However, the mean oocyte diameter of the in vitro group (69.6$\pm$2.1$\mu$m) was smaller than that of the in vivo group (73.3$\pm$3.0$\mu$m). The fertilization rate was significantly lower (p<0.05) in the in vitro group (76.5%) than in the in vivo group (90.2%), however, there was no difference in the percentage of monospermic and polyspermic oocytes between two groups. The capacities of in vitro grown ova to cleave and develop to blastocyst were (57.8 and 14.4%, respectively) significantly lower (p<0.001) than those of the in vivo counterpart (84.4 and 56.6%, respectively). Moreover, the mean number of cells per blastocyst was significantly lower (p<0.05) in the in vitro group (39.0$\pm$10.8) than in the in vivo group (60.5$\pm$12.5). Live young were produced from transferred 2-cell embryos derived from in vitro-grown and matured oocytes. In conclusion, the results show that in vitro-grown oocytes did not achieve the developmental capacity of in vitro-grown oocytes.