• Reproductive and Developmental Biology
• /
• v.29 no.4
• /
• pp.241-245
• /
• 2005

Porcine oviduct epithelial cells (POEC) are widely used in co-culture experiments to improve early embryonic development, in vitro fertilization in embryo transfer programs for domestic animals and in vitro maturation of immature germ cells. POEC were mechanically isolated and cultured in tissue culture medium 199. Cells grew continuously, and confluent monolayers were formed after 7 days. After forming confluent monolayer of epithelial cells, supernatant was collected as the condition medium for maturing round spermatids in vitro. Round spermatids were also separated mechanically and cultured in the POEC condition medium. In this study we observed that $20\%$ of round spermatid cultured were matured into elongating spermatid after 24 h, and about $10\%$ of round spermatid cultured showed complete elongation (elongated spermatid) within $24\~48$ h of in vitro culture. No further development was observed within $50\~72$ h and transformed cells lost their viability after 72 h. These preliminary findings suggest that the condition medium from POEC may be possible to overcome the round spermatid block by improving the milieu of culture system.

• Development and Reproduction
• /
• v.18 no.3
• /
• pp.153-160
• /
• 2014

Adaptive development of early stage embryo is well established and recently it is explored that the mammalian embryos also have adaptive ability to the stressful environment. However, the mechanisms are largely unknown. In this study, to evaluate the possible role of aquaporin in early embryo developmental adaptation, the expression of aquaporin (AQP) 5 gene which is detected during early development were examined by the environmental condition. To compare expression patterns between in vivo and in vitro, we conducted quantitative RT-PCR and analyzed localization of the AQP5 by whole mount immunofluorescence. At in vivo condition, Aqp5 expressed in oocyte and in all the stages of preimplantation embryo. It showed peak at 2-cell stage and decreased continuously until morula stage. At in vitro condition, Aqp5 expression pattern was similar with in vivo embryos. It expressed both at embryonic genome activation phase and second mid-preimplantation gene activation phase, but the fold changes were modified between in vivo embryos and in vitro embryos. During in vivo development, AQP5 was mainly localized in apical membrane of blastomeres of 4-cell and 8-cell stage embryos, and then it was localized in cytoplasm. However, the main localization area of AQP5 was dramatically shifted after 8-cell stage from cytoplasm to nucleus by in vitro development. Those results explore the modification of Aqp5 expression levels and location of its final products by in vitro culture. It suggests that expression of Aqp5 and the roles of AQP5 in homeostasis can be modulated by in vitro culture, and that early stage embryos can develop successfully by themselves adapting to their condition through modulation of the specific gene expression and localization.

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

• Journal of Embryo Transfer
• /
• v.5 no.2
• /
• pp.45-55
• /
• 1990

The in-vitro fertilization in human clinic and animal reproduction is a very important technique but the rate of success is still low. When the in-vitro fertilization and culture of gametes or embryos were done under the condition which Hema hydrogel chamber were implanted into the peritoneal cavity of mouse, the in-vitro fertilization and development of embryos could be significantly improved and the cell-block under in-vitro culture could be overcome. Also, the Rema hydrogel chamber was very useful for the protection of isolated blastomeres. It is concluded that the polymerized Hema (pHema) hydrogel chamber may be effectively used in the fields of embryo transfer and in vitro fertilization.

• Journal of Bio-Environment Control
• /
• v.18 no.1
• /
• pp.23-28
• /
• 2009

This study was conducted to evaluate the effect of optimized environment conditions with growth stage in photoautotrophic micropropagation on the growth of potato (Solanum tuberosum L. cv. Dejima) plantlets and energy efficiency. Optimum environment conditions at each stage were decided in our previous study. For the evaluation of optimized environment control, potato plantlets were cultured under four different conditions: photoautotrophic optimum conditions of photosynthetic photon flux density (PPFD) and $CO_2$ levels with growth stage (POG), photoautotrophic constant condition with average PPFD and $CO_2$ levels (PCA), photoauototrophic constant condition with maximum PPFD and $CO_2$ levels (PCM), and photomixotrophic conventional condition with 3% sucrose (PMC) as control. As a result, environment control with growth stage (POG) significantly promoted all the growth characteristics such as the number of nodes and unfolded leaves, shoot height, shoot diameter, and fresh and dry weights of potato grown in vitro. In addition, based on dry weight consumed electricity and $CO_2$ were the lowest in POG suggesting the highest energy efficiency among the treatments. After transferring potato plantlets to greenhouse, the plantlets under POG showed vigorous growth, which was pretty similar with those under PMC. The accumulations of dry matter in POG were 4.7 times in vitro and 3.8 times in greenhouse as much as those in the conventional control (PCM). Thus, we concluded that in vitro environment control with growth stage induced vigorous growth of potato plantlets both in vitro and in greenhouse with less energy consumption.

• Korean Journal of Plant Resources
• /
• v.11 no.1
• /
• pp.106-110
• /
• 1998

The present study has been undertake to obtain the fundatmental data of optimum germination condition and to establish storage time for artificial pollinationin Codonopsis lancelata pollen. In vitro condition for germination of freshly collected and stored pollen were examined. The optium temperature for germination of fresh pollen was $25^{\circ}C$. The optium sucrose concentration in the medium ranged from 30 to 40 % and optium pH 6.0% for pollen germination. The rate of pollen germination accelerated considerably in the medium with 1% agar. 30% sucrose, and pH 6. C. lanceolata pollen remained viable for 15 days when stored at 5$^{\circ}C$ with silica gel as desiccant.

• Journal of the Korean Society of Tobacco Science
• /
• v.30 no.1
• /
• pp.14-24
• /
• 2008

The objective of this study was to determine the effect of smoking conditions on the in vitro toxicological activity of mainstream smoke. The 2R4F reference cigarette was machine-smoked by International Organization for Standardization(ISO) and Canadian Intense(CI) conditions. Smoke was analysed for chemical composition and in vitro toxicity. The cytotoxic potencies of both the total particulate matter(TPM), which were collected in Cambridge filter pad, and gas/vapor phase(GVP), which was bubbled through in phosphate-buffer saline in a gas-washing bottle, were assessed neutral red up take assay with chinese hamster ovary(CHO) cells. The assessment for genotoxicity of TPMs generated under ISO and CI conditions was determined using Salmonella mutagenicity assay and in vitro micronucleus assay. When calculated on an equal TPM basis, in vitro toxicity of TPM obtained under CI condition was decreased compared to TPM generated under ISO condition. The results of chemical composition analyses revealed that the lower toxicological activity under CI condition than that of ISO condition could be explained by the decreased in the contents of phenols, N-nitrosoamines and aromatic amines of TPM on an equal TPM basis.

• Korean Journal of Plant Resources
• /
• v.27 no.3
• /
• pp.256-262
• /
• 2014

Viscum album var. coloratum (mistletoe) is considered as one of the endangered plant species in Korea. Our objective is to restore its population and multiplication of plant by ex situ method. In this research we explored the maximum germination (in vitro) of freshly harvested and stored seeds of mistletoe collected in different time intervals. Ex vitro germination after artificial inoculation on the branches of Prunus mume in different physiological conditions was also monitored. The research revealed that the lately harvested seeds (Feb. and March 2014) were superior over early harvested seeds (Nov. 2013 and Jan. 2014) of mistletoe due to the higher percentage of germination (above 93%). According to the data, it is also revealed that the survival and germination rate of mistletoe seeds decreased with the increase in storage duration. In ex vitro germination, the fluctuated temperature of a glass house in natural condition enhanced (four fold) the rate of germination on the branches of Prunus mume than the constant temperature condition in the glass house.

• Journal of Species Research
• /
• v.6 no.1
• /
• pp.68-75
• /
• 2017

We investigated the effects of sodium hypochlorite (NaOCl) and culture medium on embryo swelling and germination of Calanthe discolor Lindl., and established a method for determining the swelling and protocorm formation of C. discolor seeds via in vitro examination of immature seeds. Treatment of immature seeds with NaOCl greatly enhanced the extent of embryo swelling and protocorm formation of immature zygote embryos compared to seeds without NaOCl treatment. The effects of the culture media were also evaluated with regard to embryo swelling and protocorm formation of in vitro cultured seeds with and without NaOCl treatment. Additionally, the effects of white fluorescent light and red and blue LEDs lights on seedling growth in in vitro culture were examined. The most suitable condition for seedling growth after 12 weeks of culture was the red LEDs light with POM medium. These results show effective asymbiotic germination and growth of C. discolor.

• Journal of Pharmaceutical Investigation
• /
• v.32 no.4
• /
• pp.321-325
• /
• 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).