• Korean Journal of Animal Reproduction
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• v.22 no.2
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• pp.111-117
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• 1998

This study was conducted to examine the effect of culture supernatant of bovine oviductal epithelial cell(BOEC) on in vitro development of mouse embryos. To obtain the culture supernatant, ampullary epithelial cell, ithmic epithelial cell and ciliated eptithelial cell of bovine oviduct were cultured in Ham's F-10 su, pp.emented with 10% FCS. The development rates of mouse embryos to blastocyst stage were significantly(P<0.05) higher in BOEC-culture supernatant(72.3∼82.3%) than in Ham's F-10(50.7%). The proportions of embryonic development into hatched blastocysts were significantly(P<0.05) higher in ampullary cell supernatant(43.2%), ithmic cell supernatant(48.4%) and ciliated cell supernatant (27.7%) than in Ham's F-10(14.4%). On the other hand, the effect of ciliated cell supernatant was lower than those of other cell supernatants(P<0.05). And there was no difference between ampullary cell supernatant and ithmic cell supernatnat.

• Journal of Veterinary Clinics
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• v.16 no.1
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• pp.31-36
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• 1999

The aim of this study is to determine the phagocytosis-promoting factor(s) for feline peripheral blood polymorphonuclear cells (PMN) by culture supernatant from mono-nuclear cells (MNC) treated with egg white derivatives (EWD). The phagocytic activity of PMN was analyzed by a flow cytometry system. The EWD did not show direct effect on the phagocytic response of PMN. The phagocytic activity of PMN was enhanced by culture supernatant from MNC but not PMN treated with EWD. Therefore, it was suggested that the enhanced phagocytic activity of feline PMN could be mediated by humoral factor(s) released from MNC treated with EWD. Thus, the phagocytosis-promoting factor(s) in supernatant fraction from MNC culture treated with EWD were isolated by reverse phase high pressure liquid chromatography. The resulting supernatant fraction on 29.02 minutes of retention time showed high phagocytic activity of PMN. The molecular weight of this supernatant fraction was 16 to 18 kDa when analyzed by capillary electrophoresis. The isoelectric point was pH 5.76 when assessed by ion-exchange chromatography. These results suggest that EWD stimulates feline MNC to elaborate a phagocytosis-promoting factor, 16 to 18 kDa of molecular weight, which could be an important mediator for the enhancement of phagocytic activity of feline peripheral blood phagocytes. Further study will be needed to elucidate this phagocytic factor.

• Journal of Life Science
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• v.19 no.12
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• pp.1764-1768
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• 2009

This study was carried out to examine a concentration of steroid hormones and in vitro development of mouse embryos in culture supernatant of bovine granulosa cells (GC). To obtain the culture supernatant, granulosa cells were retrieved from mature follicles (6~15 mm diameter) and immature follicles (2~5 mm diameter) of bovine ovary and were cultured, respectively, in media of Ham's F-10 with 15% FCS for 16 days. Mature and immature granulosa cells formed their monolayers easily and showed similar growth patterns in culturing. There was no morphological difference between mature and immature granulosa cells. High levels of both progesterone and estradiol were detected in the culture supernatant of mature granulosa cells and immature granulsa cells, and the endocrine profiles of the two types of cells were similar. Progesterone secretion of granulosa cells was high in the late stage of culturing and estradiol secretion was high in the early stage of culturing. In vitro development rates of mouse embryos to morula, blastocyst and hatched blastocyst were significantly (p<0.05) higher in culture supernatant of mature granulosa cells (92.7%, 78.1% and 34.5%) and in culture supernatant of immature granulosa cells (96.4%, 78.5% and 26.8%) than in Ham's F-10 (86.7%, 41,7% and 13.3%). However, there was no difference between the culture supernatant of mature granulosa cells and the culture supernatant of immature granulosa cells in the development of embryos.

• Journal of Microbiology and Biotechnology
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• v.12 no.3
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• pp.526-529
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• 2002

Lysis of erythrocytes isolated from human, rabbit, and mouse blood samples was investigated with the culture supernatant of Xenorhabdus nematophilus in a primary form. Prior to use, the culture supernatant of the bacteria was concentrated and the concentrate was dialyzed against Tris-HCl buffer (10 mM, pH 8.1) by ultrafiltration using PM-5 membrane with a molecular weight cut-off of 5,000. At $30^{\circ}C$, the supernatant exhibited no lytic activity towards three types of erythrocytes. However, at $4^{\circ}C$, the supernatant showed selective lytic activity towards rabbit erythrocytes within 90 min. yet did not lyze human or mouse erythrocytes. Microscopic examination clearly revealed that most of the rabbit erythrocytes had been fumed into ghost forms.

• Microbiology and Biotechnology Letters
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• v.21 no.2
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• pp.144-149
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• 1993

We have identified a T cell-activating material in the culture supernatant of Streptomyces species. The factor in microbial culture supernatant (MCS) induced thymocyte proliferation in a does dependent fashion and it could be detected by immunoblot analysis using anti-interleukin-1(IL-1) antibody. The factor in MCS was slightly larger(about 21 kd) in its molecular weight than IL-1 on SDS-PAGE. When 125I-MCS was covalently coupled with homo-bifunctional cross-linking agent, disuccinimidyl-propionate to IL-1 receptor(IL-1R) on mouse thymoma cell(EL-4) and immunoprecipitated with anti-IL-1R antibody the molecular weight of this complex of 110 kd was observed.

• Journal of Veterinary Clinics
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• v.37 no.2
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• pp.61-66
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• 2020

Nickel is a nutritionally essential trace element that plays an important role in the immune system of several animal species. The aim of this study was to examine the effect of nickel chloride on chemotactic activity of peripheral blood polymorphonuclear cells (PMNs) and whether this effect is associated with interleukin (IL)-8 and a nuclear factor-kappa B (NF-κB)-dependent pathway. Peripheral blood mononuclear cells (PBMCs) and PMNs were isolated by Percoll solution (Specific gravity; 1.080) and 1.5% dextran treatment, respectively. A modified Boyden chamber assay was used to measure the chemotactic activity of PMNs. The level of IL-8 in culture supernatant from PBMCs was measured by enzyme-linked immunosorbent assay (ELISA). Both of PBMCs and PMNs exhibited a low viability when cultured with concentration of greater than 1,000 μM of nickel chloride for 24 h. Thus, nickel chloride was used at concentration of 500 μM, which preserved cell viability. Treatment with nickel did not directly affect the chemotactic activity of PMNs. However, the chemotactic activity of PMNs was remarkably increased by culture supernatant from PBMCs treated with nickel chloride (500 μM) for 24 h. Recombinant porcine IL-8 polyclonal antibody (pAb) neutralized the enhancing effect on the chemotactic activity of PMNs by culture supernatant from PBMCs treated with nickel and this culture supernatant had higher IL-8 levels than the culture supernatant from untreated PBMCs. In addition, n-tosyll-phenylalanine chloromethyl ketone (TPCK), a NF-κB inhibitor, antagonized the enhancing effect on the chemotactic activity of PMNs by the culture supernatant from PBMCs treated with nickel. These results suggested that nickel stimulates porcine PBMCs to produce IL-8, which increases the chemotaxis of PMNs via NF-κB-dependent pathway.

• Journal of Microbiology and Biotechnology
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• v.21 no.6
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• pp.604-612
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• 2011

Beauveria bassiana SFB-205 supernatant can effectively control cotton aphid populations, which is closely associated with its chitinase activity. The present work extends to optimizing a culture medium to produce more efficacious supernatant in flask conditions, followed by scale-up in 7 L, 300 L and 1.2 KL fermentors with the parameter of chitinase. In flask conditions, a combination of soluble starch and yeast extract produced the greatest amount of chitinase (5.1 units/ml) and its supernatant had the highest aphicidal activity. An optimal quantitative combination of the two substrates, estimated by a response surface method, enabled the supernatant to have 15.7 units/ml of chitinase activity and 3.7 ml/l of median lethal concentration ($LC_{50}$) of toxicity against cotton aphid adults in laboratory conditions. In the scale-up conditions, overall supernatant had 25-28 units/ml of chitinase activity. Decrease in pH and limitation of dissolved oxygen (DO) during cultures were significantly related to the yield of chitinase. These results suggest that the substrate-dependent chitinase production can be background information for optimizing a culture medium, and pH and DO are critical factors in maximizing the production in scale-up conditions.

• Journal of Veterinary Clinics
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• v.35 no.5
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• pp.195-199
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• 2018

Zinc is necessary for normal functions in the immune system. The objective of the study is to examine the effect of zinc on the chemotactic activity of porcine peripheral blood polymorphonuclear cells (PMNs). A modified Boyden chamber was used to determine the directional migration distance of PMNs. Various concentrations of zinc showed no chemotactic activity to PMNs. However, culture supernatant from peripheral blood mononuclear cells (PBMCs) treated with zinc remarkably increased the chemotactic activity of PMNs when compared with culture supernatant from PBMCs treated without zinc. Culture supernatant from PBMCs treated without zinc also increased the migration distance of PMNs relative to vehicle control (medium alone). Increasing effect in chemotactic activity of PMNs by culture supernatant from PBMCs treated with zinc was inhibited by treatment of porcine anti-interleukin (IL)-8 polyclonal antibody (pAb). This effect was not affected by heat treatment ($4-85^{\circ}C$). This corresponded with heat stable physical characteristics of IL-8. These results suggest that zinc can upregulate the chemotaxis of PMNs, which is primary mediated by IL-8 chemotactic factor released from PBMCs treated with zinc.

• Toxicological Research
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• v.14 no.3
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• pp.301-306
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• 1998

This experiment was carrid out to know the effects of heating and serum on hydroxyl radicals in embryonic mouse forebrain (cerebrum) culture. The heating to mouse embryonic cerebrum cells in culture was done in a water bath at 43${\circ}C$ for 60min. After that, two supernatants were prepared at 20 hrs and 48 hrs respectively after heat treatment to the brain cells. To find out the heating effects on neuron cells, mouse cerebrum cells (13 embryonic day) were cultured in hydroxyl radical generation system composed of 20mU/ml glucose oxidase (GO system), using condition of normal culture media (MEM, 5% serum, 5% $CO_2$or supernatant prepared after heating at 43${\circ}C$ for 60 min in a water bath. Supernatant prepared at 20 hrs after heat treatment had a greater protective effects against hydroxyl radical than supernatant prepared at 48 hrs after heat treatment . Otherwise, the protective effect of serum against hydroxyl radicals in the cultured brain cells is higher than that in the heat treatment. These results indicated that serum in culture media reduced cytotoxicity of hydroxyl radicals in mouse forebrain culture, also that heat treatment showed the protective effects against hydroxyl radicals generated with 20mU/ml GO system in mouse forebrain culture.

• Restorative Dentistry and Endodontics
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• v.22 no.2
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• pp.761-768
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• 1997

Microorganisms and their by-products are considered to be the major causes of pulpal and periapical pathosis. The role of microorganisms in endodotic infection has been studied regarding the prevalence of particular organisms found in root canal and periapical lesions. The aim of this study was to investigate the effect of culture supernatants of several oral microorganims isolated from infected root canals on the viability of cultured cell lines using colorimetirc assay. S. simulans, S. sciuri, E. faecium, S. intermedius, S. mitis, S. sanguis and S. uberis were incubated in Todd-Hewitt broth for 16 hours. 20 and 100ul of filtered bacterial cell culture supernatants were added to MK and Hep-2 cells. Cell viability was measured using MIT colorimetric assay. 20ul and 100u1 of S. sanguis supernatant showed significant cytotoxicity compared to control on MK cells. 100ul of S. sanguis supernatant significantly depressed viability of HEp-2 cells. E. faecium and S. intermedius did not affect the viability of MK and HEp-2 cells.