• Proceedings of the Plant Resources Society of Korea Conference
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• 2002.04a
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• pp.43-44
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• 2002

• Korean Journal of Animal Reproduction
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• v.16 no.4
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• pp.341-346
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• 1993

The development of isolated blastomeres from mammalian preimplantation embryos has been basically studied for the multiplication of embryos from superior animals. Therefore, this study was investigated the effect of co-culture with mouse fetal fibroblast cells(MFFC) on in vitro development of blastomeres from mouse preimplantation embryos. Mature female ICR mice were treated with hormone to induce superovulation and embryos were collected at each 2, 4, and 8-cell stage. Then, after removing zona pellucida with protease, blastomeres were isolated by micropipetting, or reconstituted with different stage blastomere, and incubated for 72 hrs either in T6 or TCM199 or on the monolayer of MFFC, which was prepared with fibroblast cells from 14∼14 day mouse fetus. After incubation, we examined their development rates every day and the nuclei numbers of each blastocyst by Hoechst-33342 staining. In the development rates of blastomeres, there were no significant differences between media but the higher rateswere found in the monolayer of MFFC, regardless of reconsititution. In addition, blastomeres cultured with MFFC had slightly greater number of nuclei than those cultured in single media. Generally, the higher development rates of blastomeres were found from earlier stage embryos than the later ones, regardless of culture conditions. Reconsitituted blastomeres had more nuclei but did not show the higher development rates, compared to the single blastomeres. Taken together, our results suggest that co-culture with MFFC have a beneficial effect on the in vitro development of blastomeres from mouse embryos.

• Journal of Animal Science and Technology
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• v.58 no.3
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• pp.11.1-11.6
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• 2016

Background: This study was investigated the effects of culture media conditions on production of eggs fertilized in vitro of embryos from ovaries of high grade Korean native cow, Hanwoo. Methods: The IVMD 101 and IVF 100 were used for in vitro maturation of selected Hanwoo oocytes and In vitro embryo culture after in vitro fertilization, respectively. The IVMD 101 and IVD 101 were used for in vitro culture and completely free of serum. Results: The cleavage rates of 2-cell embryos in reference to Hanwoo oocytes were 86.7, 92.9, and 90.1 % in the control group, IVDM101 medium and IVD101 medium, respectively which indicates that the IVDM101 medium and IVD101 medium may result favorable outcomes. The in vitro development rates of blastocysts were 12.4, 38.4 and 32.4 % in the control group, serum free IVMD101 medium and IVD101 medium, respectively. For hatched blastocysts, it was 5.3, 33.9, and 28.6 % in the control group, serum free IVMD101 medium and IVD101 medium, respectively. Hence, more favorable results were expected for the hatched blastocysts in which the IVMD101 medium and IVD101 medium were used than the control group. Average cell numbers of blastocysts were 128.3, 165.7, and 163.6 in the groups of TCM-199 + 10 % FBS medium, IVMD 101 medium, and IVD 101 medium, respectively which clearly show that the IVMD 101 and IVD 101 medium consequence significantly higher cell numbers compared to the control group (i.e., TCM-199 +10 % FBS medium). Pregnancy rate after embryo transfer was 39.6 % when the serum free medium was used which is higher than that of the medium supplemented with serum (32.8 %). In addition, stillbirth rates were 4.9 % in the group of serum free medium whereas it was 13.6 % in the serum supplemented medium (13.6 %). Conclusions: Taken altogether, serum free media, the IVMD 101 and IVD 101 represented more favorable results in the embryo development rate of embryos, cell numbers of blastocyst, and pregnancy rate. Of note, the IVMD 101 medium showed better outcomes hence, it might be a better option for future applications for in vitro culture of bovine embryos.

• Clinical and Experimental Reproductive Medicine
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• v.49 no.4
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• pp.225-238
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• 2022

The ultimate goal of human assisted reproductive technology is to achieve a healthy pregnancy and birth, ideally from the selection and transfer of a single competent embryo. Recently, techniques for efficiently evaluating the state and quality of preimplantation embryos using time-lapse imaging systems have been applied. Artificial intelligence programs based on deep learning technology and big data analysis of time-lapse monitoring system during in vitro culture of preimplantation embryos have also been rapidly developed. In addition, several molecular markers of the secretome have been successfully analyzed in spent embryo culture media, which could easily be obtained during in vitro embryo culture. It is also possible to analyze small amounts of cell-free nucleic acids, mitochondrial nucleic acids, miRNA, and long non-coding RNA derived from embryos using real-time polymerase chain reaction (PCR) or digital PCR, as well as next-generation sequencing. Various efforts are being made to use non-invasive evaluation of embryo quality (NiEEQ) to select the embryo with the best developmental competence. However, each NiEEQ method has some limitations that should be evaluated case by case. Therefore, an integrated analysis strategy fusing several NiEEQ methods should be urgently developed and confirmed by proper clinical trials.

• Korean Journal of Medicinal Crop Science
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• v.20 no.5
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• pp.301-307
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• 2012

IPP isomerase (Iso) and Limonene synthase (Limo) are important enzymes in terpenoids biosynthesis pathway. The wild type and each metabolically engineered (Iso and Limo) transgenic spearmint (Mentha spicata Linne) plants were compared for their growth patterns and the contents of essential oil in in vitro culture media. The profile of terpenoid metabolites was obtained from the essential oil of the metabolically engineered transgenic spearmint, which was extracted using a modified SDE method, by GC-MS analysis. The growth of wild spearmint was more profuse in B5 culture medium than in other media. Significant differences in leaf and root growth patterns were observed between metabolically engineered transgenic and wild type spearmint plants. The leaves of the transgenic spearmint plants were slightly elongated but were dramatically narrower than those of wild type spearmints. The content of essential oil of transgenic spearmint was different slightly depending on the target terpenoid genes. The content of essential oils in Limo transgenic plants was higher than that of Iso, except for transgenic plant in B5 medium. The transgenic spearmint produced more terpenoids than the wild type. Iso spearmint extracts showed eleven terpenoids and a phenylpropane, while Limo spearmint extracts contained nine terpenoids. However, extracts from the wild type showed the presence of only four terpenoids.

• Clinical and Experimental Reproductive Medicine
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• v.16 no.2
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• pp.161-171
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• 1989

The development of 2-cell mouse embryos to the blastocyst stage in vitro has been used as quality control for the culture media and supplements employed for human in vitro fertilization and embryo transfer(IVF-ET). 2-cell mouse embryos were cultured to the blastocyst stage in SECM, Medium 199-Earle's, Ham's F-10 I , Ham's F-10 II , Hoppe & Pitts, MEM and $HT_6$. The protein supplements contained in media were bovine serum albumine, fetal bovine serum and human fetal cord serum. The results were as follows; 1. The successful development was 81.3% in Medium 199-Earle’s, 91.9% in Ham’s F-10 I and 97.1% in $HT_6$. 2. 2-cell mouse embryos developed properly in all supplements but the best development was particularly noted in $HT_6$ media when HFCS was supplied as protein supplement.

• Journal of Plant Biotechnology
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• v.30 no.3
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• pp.269-273
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• 2003

In order to develop an efficient micropropagation protocal for Eucalyptus pellita, on in vitro culture system has been was established by inducing axillary buds from greenhouse stock materials. Among 6different media tested, DKW medium was the best ot induce bast induce both shoot proliferation and growth. Average number of proliferated shoots of 403per explant was obtained at the concentration of 0.1mg/LBA. Most of the stem materials excreted phenolic compounds at the proximal part of the explant and caused darking of the media. Therefore, it was necessary to transfer frequently to a fresh medium and/or to add activated charcoal at the concentration of 0.02％(w/v). Generally on vitro roots were formed easily on 1/2DKW medium with NAA treatment. All the explants rooted at the medium containing 0.2mg/L NAA and displayed vigorous root growth in vitro culture conditions. After transferred to an artificial soil mixture (peatmoss: vermiculrite: perlite, 1:1:1, v/v/v) in the greenhouse, most rooted plantlets survived well without any morphological abnormalities. The results show that the species can be micropropagated effectively by the application of axillary bud culture system.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2000.04a
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• pp.201-207
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• 2000

The strategies of commercial development have been focused on the economy of scale for a process. The design of media has been recognized as a key in assuring mass production of entomopathogenic nematodes. Media optimization was conducted with insect host, proteins, lipids, and symbiotic bacteria mass. G. mellonella (insect host) produced about 290,000 infective juveniles per one. Complex media produced about 250,000 infective juveniles / ml in liquid culture within 8 days (one generation).

• Journal of Pharmaceutical Investigation
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• v.40 no.6
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• pp.363-366
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• 2010

Identification of compounds that function as P-glycoprotein (P-gp) substrates or inhibitors can facilitate the selection and optimization of new drug candidates. The purpose of this study is to optimize the experimental conditions for in vitro P-gp assay using LLC-GA5 cells, which is a well-known transformant cell line derived by transfecting LLC-PK1 with human MDR1. The amount of rhodamine123 transported by the LLC-GA5 and LLC-PK1 cells was evaluated under the following experimental conditions: 3 different types of transport media, colchicine pretreatment or nontreatment of the cells in the culture media, and with and without poly-L-lysine coating of the culture plates. The assay sensitivity was found to considerably differ depending on the diluents used in the transport media. P-gp-mediated transport in LLC-GA5 cells was most clearly characterized in the Hanks' balanced salt solution based transport media. The sensitivity of P-gp-mediated transport was not changed by colchicine pretreatment or poly-L-lysine coating of the culture plates.

• Korean Journal of Animal Reproduction
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• v.14 no.1
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• pp.50-56
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• 1990

Bovine oocytes obtained from follicles(2~5mm) of ovaries after slaughter were cultured in TCM 199 medium with 10~20% heat-inactivated estrus cow serum(ECS) for 25~27 hr, at 39$^{\circ}C$ under 5% CO2 in air. At the end of culture period, some oocytes were stained with 1% acetoorcein and examined for the evidence of oocyte maturation. The remainder were used to assess the potential of in vitro fertilization(IVF) with frozen-thawed spermatozoa and subsequent development in media with or without bovine oviduct epithelial cell (BOEC) co-culture. The results obtained were summarized as follows ; 1. The maturation rate of oocyte in vitro in TCM 199 medium with 15% ECS group(76.3) was superior to 10% ECS group(68.3%) and 20% ECS group(64.5%). 2. The IVF rates of oocytes matured in vitro, and formation rate of male and female pronuclei were 63.6%(77/121) and 93.5%(72/77), respectively. The incidence of polyspermy was very low(2.4%). 3. Of 73 oocytes fertilized in vitro and cultured in TCM 199 medium with 10% fetal calf serum for 7 days, 41(56.3%) were cleaved over 2-cell and only 1(2.4%) was developed beyond the 16-cell stage. 4. Of 76 oocytes co-cultured with BOEC, 58(76.3%) were cleavaged and 23(39.7%) were developed to morula and blastocyst stage. The results of this study indicate that co-culture with BOEC deserved a positive effect on the IVF oocyte development through the 16-cell block.