• Journal of Plant Biology
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• v.38 no.2
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• pp.143-151
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• 1995

Addition of plant growth hormones [0.01 mg/L NAA and 0.2mg/L benzyladenine (BA)] to a woody plant medium stimulated the adventitious bud formation of poplar explants during culture. Endogenous IAA content increased rapidly at the initial culture stage and then decreased, being followed by rapid increment again at the late culture. But the content of trans-zeatin riboside (t-ZR) increased continuously during the culture. Cytoplasmic soluble proteins were analyzed by one- and two-dimensional SDS-PAGE. Increased amount of 40 kD band was detected by one-dimensional electrophoresis using Coomassie Blue staining during the culture and two distinctive proteins whose mol wt is 40,000 were detected by two-dimensional electrophoresis using autoradiography and these proteins were synthesized continuously prior to the adventitious bud formation. When the midvein segments were transferred to the actinomycin D-containing medium, the spots of adventitious bud-related proteins(ABRPs) did not disappeared but weakened in intensity. So, it is concluded that genes coding for the ABRPs are regulated to some degree at the transcriptional level. Also, they were not observed in BA-free medium, suggesting that these proteins be regulated by cytokinin, which made then possible to form the adventitious bud.

• Clinical and Experimental Reproductive Medicine
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• v.41 no.1
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• pp.1-8
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• 2014

Objective: Estrogen related receptor ${\beta}$ (Esrrb) is a member of the orphan nuclear receptors and may regulate the expression of pluripotencyrelated genes, such as Oct4 and Nanog. Therefore, in the present study, we have developed a method for delivering exogenous ESRRB recombinant protein into embryos by using cell-penetrating peptide (CPP) conjugation and have analyzed their effect on embryonic development. Methods: Mouse oocytes and embryos were obtained from superovulated mice. The expression of Oct4 mRNA and the cell number of inner cell mass (ICM) in the in vitro-derived and in vivo-derived blastocysts were first analyzed by real time-reverse transcription-polymerase chain reaction and differential staining. Then 8-cell embryos were cultured in KSOM media with or without $2{\mu}g/mL$ CPP-ESRRB protein for 24 to 48 hours, followed by checking their integration into embryos during in vitro culture by Western blot and immunocytochemistry. Results: Expression of Oct4 and the cell number of ICM were lower in the in vitro-derived blastocysts than in the in vivo-derived ones (p<0.05). In the blastocysts derived from the CPP-ESRRB-treated group, expression of Oct4 was greater than in the non-treated groups (p<0.05). Although no difference in embryonic development was observed between the treated and non-treated groups, the cell number of ICM was greater in the CPP-ESRRB-treated group. Conclusion: Treatment of CPP-ESRRB during cultivation could increase embryos' expression of Oct4 and the formation rate of the ICM in the blastocyst. Additionally, an exogenous delivery system of CPP-conjugated protein would be a useful tool for improving embryo culture systems.

• Proceedings of the KSAR Conference
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• 2002.06a
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• pp.97-97
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• 2002

Achieving of in vitro development for mammalian premature spermatogenic cells are very difficult. In-vitro culture of spermatogenic cells were then initiated in an effort to try to study in vivo spermatogenesis and to understand its molecular events. Recently, the morphogenetic changes of spermatocytes or spermatid by in-vitro culture system were achieved. (omitted)

• Journal of Sericultural and Entomological Science
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• v.36 no.2
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• pp.119-123
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• 1994

By in vivo labelling of AMHP using[35S]-methionine, fat body culture and immunological analysis, it is proved that Bombyx adult fat body synthesizes 18K and 20K subunits of AMHP and releases them into haemolymph. Also these peptides are assembled to form native AMHP in the adult haemolymph.

• Korean Journal of Animal Reproduction
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• v.16 no.4
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• pp.347-352
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• 1993

This study was designed to develop the in vitro culture systemof mammalian preimplantation embryos. We proposed mouse fetal fibroblast cells (MFFC) from 14∼15 day mouse fetus. Zygotes from superovulated female ICR mice were cultured 96 hrs in simple defined media (T6) or on the monolayer of MFFC. In addition, to evaluate the action of the co-culture of MFFC, various diluted superoxide dismutase antibody (SOD-Ab) was supplemented into the monolayer of MFFC and zygotes were cultrued in presence or absence of SOD-Ab. The developmental rates of zygotes were significantly increased in co-culture with MFFC compared to the control. The rates of zygotes to the 4-cell stage in media treated with EDTA were higher than those cultured in MFFC but the proportions of morula and blastocyst were not differ between EDTA and MFFC. Interestingly blastocysts in co-culture with MFFC possessed as many as blastomere as those developing in vivo, but blastocysts cultured with EDTA had significantly fewer blastomeres. In addition, the treatment of SOD-Ab suppressed the beneficial effect of MFFC. Therefore, our findings suggest that co-cultrue system using MFFC may have an advantage in the development of mouse zygotes as well as embryonic differentiation.

• Proceedings of the KACD Conference
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• 2003.11a
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• pp.583-583
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• 2003

Recent study reported whether the cultured human pulp cells increase IL-8 secretion in response to SP stimulation22). In the present study, whether induction of IL-8 or MCP-1 in pulp tissue can be detected using enzyme-linked immunosorbent assay(ELISA) with ex vivo pulpal explants exposed to neuropeptides in culture and the IL-8 expression using immunohistochemical analysis with the ex vivo pulpal explants exposed to neuropeptides was evaluated. To investigate further mechanisms that may contribute to leukocyte recruitment in lesions of endodontic origin, the differential expression of IL-8 and MCP-1 by human dental pulp tissues stimulated in vitro by the Substance P was examined.(omitted)

• Journal of Embryo Transfer
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• v.30 no.4
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• pp.315-317
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• 2015

In vitro culture of murine embryos is an important step for in vitro production systems including in vitro fertilization and generations of genetically engineered mice. M16 is widely used commercialized culture media for the murine embryos. Compared to other media such as potassium simplex optimization medium, commercial M16 (Sigma) media lacks of amino acid, glutamine and antibiotics. In the present study, we optimized M16 based embryo culture system using commercialized antibiotics-glutamine or amino acids supplements. In vivo derived murine zygote were M16 media were supplemented with commercial Penicillin-Streptomycin-Glutamine solution (PSG; Gibco) or MEM Non-Essential Amino Acids solution (NEAA; Gibco) as experimental design. Addition of PSG did not improved cleavage and blastocyst rates. On the other hand, cleavage rate is not different between control and NEAA treated group, however, blastocyst formation is significantly (P<0.05) improved in NEAA treated group. Developmental competence between PSG and NEAA treated groups were also compared. Between two groups, cleavage rate was similar. However, blastocyst formation rate is significantly improved in NEAA treated group. Taken together, beneficial effect of NEAA on murine embryos development was confirmed. Effect of antibiotics and glutamine addition to M16 media is still not clear in the study.

• Journal of the Korean Society of Food Science and Nutrition
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• v.13 no.4
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• pp.403-405
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• 1984

Since chlorella was found to be a source of protein in 1974, wide ranges of investigations on culture methods, its constituents and nutritional factors have been carried out, i. e. most of them were the reports on the nutritional evaluation. However, kinetics such as absorption, distribution, metabolism and elimination of chlorella protein have not been fully elucidated. So, on the assumption that using $N^{15}$ labelled chlorella protein could accomplish good results for kinetics of chlorella in in vivo experiments, $N^{15}$ was added to the culture fluid. From the result of this study, it is suggested that chlorella utilizes N as well as $N^{15}$ in protein synthesis, and this $N^{15}$ labelled chlorella protein can be useful tool for the study of kinetics of chlorella in in vivo experiments.

• KSBB Journal
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• v.13 no.1
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• pp.31-37
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• 1998

A simple method for the in vivo production of third-stage infective juveniles(IJs) of Steinernema glaseri was developed. Using Galleria mellonella larvae, only IJs can be rapidly generated inadequate quantities for field application. The nematode inoculation concentration and incubation temperature were critically important. The most effective temperature for infectivity of Steinernema glaseri IJs to Galleria mellonella larvae was 33$^\circ C$. However, the total number of menatodes harvested at 25$^\circ C$ about 66,000 IJs per larva was significantly greater than those at other temperatures. The optimal inoculation number of nematodes was 60 to 80 nematodes per host larva. The higher nematode inoculation concentration of 100 IJs per larva caused a rapid decrease in the total number of IJs harvested. As the inoculation medium pH increased, the number of IJs harvested increased and reached about 110,000 IJs per larva at pH 9.0. The pathogenicity of IJs decreased y increasing the salt concentration in the medium.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.2
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• pp.67-75
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• 2002

The human complement receptor type 1 (CR 1, C3 b/C4b receptor) is a polymorphic membrane glycoprotein expressed on human erythrocytes, peripheral leukocytes, plasma and renal glomerular podocytes, which consists of transmembrane and cytoplasmic domains with 30 repeating homologous protein domains known as short consensus repeats (SCR). CR1 has been used as an inhibitor for inflammatory and immune system for the past several years. Recently; it is reported that CRl was found to suppress the hyper-acute rejection in xeno-transplantation and can be used to cure autoimmune diseases. A soluble form of CRl, called sCRl, is a recombinant CRl by cleaving the transmembrane domain at C-terminus and has been expressed in Chinese Hamster Ovary (CHO) cells. Several purification methods for sCR1 from CHO cells have been reported, but most of them require complicated steps at high cost. Moreover, such methods are mostly performed under the pH condition apt to denaturing sCR1 and causes sCRl losing its activity. We here report a rapid and efficient method to purify sCR1 from CHO cell. The new method consists of a two-stage of cell culture by cultivating cells in serum medium followed by serum-free medium, and a two-stage of column purification by means of heparin and gel filtration column chromatography. By using this novel method, sCR1 can be purified in a simple and effective way with high yield and purity, furthermore, the purified sCR1 was confirmed to retain its activity to suppress the complement activation in vivo and ex vivo.