• Journal of Animal Reproduction and Biotechnology
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• v.35 no.1
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• pp.65-72
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• 2020

Fish ovarian germline stem cells (OGSCs) that have the abilities to self-renew and differentiate into functional gametes can be used in various researches and applications. A main issue to be solved for effective utilization of fish OGSCs is the development of their stable in vitro culture condition, but only few researches about fish OGSC culture have been reported so far. In this study, in order to find the clues to develop the culture condition for OGSCs from Japanese medaka (Oryzias latipes), we tried to establish somatic cell lines as a candidate for the feeder cells and evaluated its supporting effects on the culture of ovarian cell populations from O. latipes. As the results, the somatic cell lines could be established only from the embryonic tissues among three tissues derived from embryos, fins and ovaries. Three embryonic cell lines were tested as a feeder cell for the culture of ovarian cell population and all three cell lines induced cell aggregation formation of the cultured ovarian cells whereas the feeder-free condition did not. Furthermore, a significant cellular proliferation was observed in the ovarian cells cultured on two of three cell lines. As a trial to increase the capacity of the cell lines as a feeder cell that supports the proliferation of the cultured ovarian cells, we subsequently established a stable line that expresses the foreign O. latipes fibroblast growth factor 2 (FGF2) from an embryonic cell line and evaluated its effectiveness as a feeder cell. The ovarian cells cultured on FGF2 expressing feeder cells still formed cell aggregates but did not show a significant increase in cellular proliferation compared to those cultured on non-transformed feeder cells. The results from this study will provide the fundamental information for in vitro culture of medaka OGSCs.

• Journal of Embryo Transfer
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• v.14 no.2
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• pp.113-119
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• 1999

This study was performed to improve the development of the in vitro fertilized bovine embryos by the condition of in vitro maturation. COCs were matured in TCM 199 supplemented with 0.1% PVA, 10ng/ml EGF, Hormones (5$\mu\textrm{g}$/ml FSH, 10 IU hCG, 1 $\mu\textrm{g}$/ml estradiol 17-$\beta$) or granulsa cell+Hormones atmosphere 39$^{\circ}C$, 5% CO2, 95% air for 24hrs. Matured oocytes were fertilized with frozen-thawed semen capacitated with 5mM caffein in BO medium for 20 hrs. IVF embryos were cultured in TCM 199 containing with hormones(same as matured medium), 10% FBS and co-culture with bovine oviduct epitherial cells. Maturation rates of COCs were showed 73.8%, 78.5%, 83.2% and 87.6% respectively, and were significant differences between PVA, EGF, and Hormones, GC+Hormones(p<0.05). The cleavage rates of IVF embryos were revealed 72.5%, 78.4%, 82.3% and 84.2% and showed same tendency as maturation rates(p<0.05). The blastocysts matured by above maturation condition and cultured for 7~10 days after fertilization had 34.4, 43.6, 52.3 and 59.3 cells had no differences among the treatments. These results suggest that high molecules as a substitutes of serum and growth factor may induce nuclear resumption of COCs but we need more study to produce transferable IVF blastocysts by use of that agents.

• Parasites, Hosts and Diseases
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• v.38 no.2
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• pp.59-64
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• 2000

To determine a suitable condition for in vitro infection model of cryptosporidium parvum, four different cell lines, AGS, MDCK, HCT-8 and Caco-2, were used as host cell lines which were cultured at various concentrations of added supplements. These supplement include fetal bovine serum (FBS), sodium choleate, ascorbic acid, folic acid, calcium pantothenate, para-aminobenzoic acid and pyruvate and their effects on the cell lines which were infected with C. parvum were evaluated. The results of this study showed that the AGS cell line was most susceptible to C. parvum whereas the Caco-2 cells appeared to be least susceptible to C. parvum. In regards to the serum condition, 10% FBS was suitable for the growth of AGS and HCT-8 cells, and 1% FBS was good for the growth of the MDCK cells when they were inoculated with C. parvum. Vitamines had a positive effect on the AGS cells, and pyruvate also showed positive effects on all of the cell lines except for Caco-2. Modified medium for each cell line was prepared by adding appropriate amounts of each supplement which resulted in the highest parasite infection number. Modified media increased the number of parasites infected on AGS cells to 2.3-fold higher when compared to the control media. In this study, we found that the AGS cell line was a suitable host model for evaluating C. parvum in vitro study and the media contents for the optimal infection conditions were suggested.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2002.11a
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• pp.93.2-93
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• 2002

• Proceedings of the Plant Resources Society of Korea Conference
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• 2001.11b
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• pp.111-113
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• 2001

• Proceedings of the Plant Resources Society of Korea Conference
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• 2000.05a
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• pp.80-81
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• 2000

• Journal of Technologic Dentistry
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• v.21 no.1
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• pp.145-151
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• 1999

Titanium(Ti) alloys has been mostly concerned with biocompatibility, corrosion resistance, and biofunctionality. However, very little is known, about the biological effects of titanium on microorganism and in particular on the oral flora. So, the effect of titanium on the in vitro growth of oral microorganism forming dental caries was studied under either aerobic or anaerobic condition. In this study, the mostly bacterial species commonly found in dental plaque or gingival sulcus grew well in an aqueous medium containing $100{\mu}g/ml$ of titanium standard solution.

• Korean Journal of Food Science and Technology
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• v.36 no.5
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• pp.765-772
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• 2004

Cooking condition of Tarakjuk (milk-rice porridge) was established based on gelatinization temperature using differential scanning calorimetry (DSC) of roasted Ilpum rice flour, which has highest enzyme-resistant starch (RS) content. Effect of cooking temperature and time on DSC characteristics, crystallity with X ray diffractogram, RS content, in vitro starch digestibility (IVSD), amino acid composition, and in vitro protein digestibility (IVPD) of Tarakjuk were determined. Tarakjuk was cooked at 50, 56.5, 64, and $69^{\circ}C$ for various durations. Rice flour ingredient used was Ilpum, previously roasted at $185^{\circ}C$ for 25 min. Tarakjuk cooked at 50 and $56.5^{\circ}C$ showed two thermal transitions between $63.7-125.2^{\circ}C$ as determined by DSC, corresponding to endotherms of starch gelatinization $(63.7-73.8^{\circ}C)$ and melting of amylose-lipid complex (AM-lipid complex, $97.7-125.2^{\circ}C$), whereas that cooked at 64 and $69^{\circ}C$ showed only AM-lipid complex melting transition between $96.9-127.6^{\circ}C$. As cooking temperature increased, RS content of Tarakjuk decreased, whereas IVSD increased. Total amino acid content was between 11,558-15,601mg/100g, depending on cooking condition used. Compared with conventionally made control, contents of essential amino acids, such as lysine and tryptophane, were higher at 50 and $56.5^{\circ}C$, and IVPD showed higher increase. Results reveal degree of gelatinization in Tarakjuk with high RS content as well as low IVSD and high IVPD, which are important from physiological and nutritional point of view, can be produced by controlling cooking condition.

• Korean Journal of Animal Reproduction
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• v.21 no.3
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• pp.293-302
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• 1997

Follicular oocytes of Grade I and II were collected from 2~6 mm ovarian follicles and matured in vitro (IVM) for 24 hrs in TCM-199 su, pp.emented with 35$\mu\textrm{g}$/ml FSH, 10$\mu\textrm{g}$/ml LH, and 1$\mu\textrm{g}$/ml estradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. They were fretilized in vitro (IVF) by epididymal spermatozoa capacitated with heparin for 12 hrs. The zygotes were then co-cultured in vitro with bovine oviducted epithelial cells (BOEC) for 7 to 9 days. The optimal time for IVM, the successful enucleation of IVM oocytes by micromanipulation at different oocyte ages after IVM, and the ideal culture system for IVM for effective IVF and in vitro development of IVM-IVF embryos was examined for in vitro production of nuclear recipient oocytes and nuclear donor embryos. To improve the efficiency of nuclear transplantation (NT) of IVF embryo into IVM follicular oocytes, this study evaluated the optimal electric condition and oocytes age for activation of IVM oocytes and in vitro development of NT embryos. In vitro development of NT embryos with preactivation or non-preactivation in enucleation oocytes, cell number of IVN-IVF embryos, and NT embryos wre also examined. The results obtained were as follows; 1. The most suitable enucleation time was at 24 hpm (83.3%) rather than that of 28 hpm(69.6%) and 32 hpm(50.0%). 2. There was no difference among the fusion rates of NT embryos at the voltages of 0.75, 1.0 and 1.5 kV/cm, but the in vitro development rates to morule and blastocyst were significantly (P<0.05) higher at the voltage of 0.75(12.5%) and 1.0kV/cm (12.6%) compared to 1.5kV/cm(0%). 3. No significant difference in activation rates were seen in NT embryos stimulated for 30, 60 and 120 $\mu$sec (71.7, 85.2 and 71.9%, respectively), but the in vitro development rates to morulae and blastocyst were significantly (P<0.05) higher in the oocytes stimulated for 30 $\mu$sec (11.6%) and 60 $\mu$sec(10.7%) than 120 $\mu$sec(0.0%). 4. The fusion rates (71.0 and 87.3%) and the in vitro development rates (9.1 and 12.7%) to morula and blastocyst were seen in the NT embryos stimulated at 28 and 32 hpm under the condition of 1.0 kV/ml, 60 $\mu$sec. However, at 24 hpm the fusion rates were 64.8% and the in vitro development to morula and blastocyst were not seen. 5. The fusion rates between the 8~12, 13~17 and 18~22-cell stage of IVM-IVF embryos were not significantly different. The in vitro development rates of the fused embryos to morula and blastocyst which were received from a blastomere of 8~12, 13~17 and 18~22-cell stages of IVM-IVF embryos were 14.9, 8.3 and 6.5%, respectively. 6. The in vitro development rate of the enucleated recipient oocytes with preactivation (24.2%) to morula and blastocyst was significantly (P<0.05) higher than that of non-preactivation (12.8%). 7. The cell numbers of NT blastocyst and IVM-IVF blastocyst cultured during 7~9 days were 63$\pm$11 and 119$\pm$23, and then their the mean cell cycle number were 5.98 and 6.89, respectively.

• Journal of Applied Biological Chemistry
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• v.48 no.4
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• pp.161-166
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• 2005

Among 24 isolates of fluorescent Pseudomonads, 5 isolates named as LPT1, LPT2, LPT3, LPT4 and LPT5 were screened in vitro for their nematicidal activity against cyst forming nematode, Heterodera cajani causing patchiness, poor and stunting growth besides discoloration in Sesamum indicum. Second stage juveniles of H. cajani hatched from egg masses were collected from roots of host plant and subjected to fresh and heat-treated culture filtrate of isolates for 24 h. Mortality of H. cajani was recorded on the basis of parameters used for test organism bioassay. Among these isolates, Pseudomonas aeruginosa LPT5 caused maximum mortality towards second stage juvenile of H. cajani in vitro. Five isolates were used as seed coating for the management of cyst forming nematode H. cajani on sesame in green house condition. The strains LPT5 was better than the other strains in reducing the population of H. cajani both in vitro and in vivo. The reduction in cyst and juveniles population was found to be 49 and 60%, respectively when seeds were coated with strain LPT5. Among other strains, LPT4 was also found to inhibit the cyst and juveniles population 12 and 36% respectively. Increases in early vegetative plant growth parameters recorded in both in vitro and in vivo further revealed the significance of indigenous bacteria in comparison to introduced strain.