• Journal of Veterinary Clinics
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• v.7 no.1
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• pp.419-427
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• 1990

Immature bovine oocytes were cultured to investigate whether the addition of FCS(10% or 20% ), CS (10%or 20% ) or BSA(5mg/ml) to culture medium with or without HEPES and co-culture with granulosa cells affect the frequency of in vitro maturation of extrafollicular bovine oocytes. After culture, the maturation rates were examined by the presence of 1st polar body and nuclear configuration. The maturation rate when FCS and CS as protein supplement were added to culture medium with or without HEPES was significantly higher than when BSA was added, and the maturation rate of extrafollicular bovine oocytes co-cultured with granulosa cells was higher than that cultured without granulosa cells, but there was no significant difference. FCS and CS were shown to be superior protein supplement when compared to BSA, and serum concentration, HEPES and co-culture with granulosa cells did not affect the in vitro-maturation of extrafollicular bovine oocytes.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.69-69
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• 2001

The cell cycle phase in which donor nuclei exist prior to nuclear transfer is an important factor governing developmental rates of reconstituted embryos. It was suggested that quiescent G0 and cycling G1 cells could support normal development of reconstituted embryos. In a quest of optimized donor nuclei treatment prior to nuclear transfer, this study was undertaken to examine the cell cycle characteristics of bovine fetal and adult somatic cells when cultured under a variety of culture treatments and the cell cycle change with the lapse of time after trypsinization. This was archived by measuring the DNA content of cells using flow cytometry, Cultured fetal fibroblast cells, adult skin and muscle cells, and cumulus cells were divided by 3 culture treatments; 1) grown to 60-70% confluency (cycling), 2) serum starved culture, 3) culture to confluency. Trypsinized cells were fixed by 70% ethanol and stained with propidium iodide. For one experiment, trypsinized cells were resuspended in DMEM＋10% FBS and incubated for 1.5, 3 and 6 h with occasional shaking before ethanol fixation. Cell cycle phases were determined by flow cytometry enabling calculation of percentages of G0＋G1, S and G2＋M. The majority of cells were in G0＋Gl stage regardless of origin of cells. Cultures that were serum starved or cultured to confluency contained significantly (P<0.05) higher percentages of cells in G0＋G1 (89.5-95.4%). For every cell lines and culture treatments, percentages of cells in existing in G0＋G1 increased with decreasing of the cell size from large to small. In the serum starved and confluency groups, about 98% of small cells were in G0＋G1 Serum starved culture contained higher percentages of small-sized cells (38.5-66.9%) than cycling and confluent cultures regardless of cell lines (P<0.05). After trypsinization of fetal fibroblast and adult skin cells that were serum starved and cultured to confluency, the percentages of cells in G0＋G1 significantly increased by incubation for 1.5(95.7-99.5%) and 3.0 h (95.9-98.6%). The results suggest that the efficient synchronization of bovine somatic cells in G0＋G1 for nuclear transfer can be established by incubation for a limited time period after trypsinization of serum starved or confluent cells.

• Biomedical Science Letters
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• v.14 no.1
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• pp.33-38
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• 2008

In order to examine oxidative stress of reactive oxygen species and the antioxidant effect of Citri Reticulatae Pericarpium (CRP) extract, human skin melanoma cells were treated with various concentrations of hydrogen peroxide ($H_2O_2$). Antioxidant effect of CRP extract on $H_2O_2$-induced cytotoxicity, cell viability, DPPH-radical scavenging activity and superoxide dismutase (SOD)-like activity. In this study, $H_2O_2$ decreased cell viability of cultured human skin melanoma cells in dose- and time-dependent manners, and then, midcytotoxicity value (MCV) was determined at $60\;{\mu}M$ after human skin melanoma cells were cultured for 5 hours in the media containing $20{\sim}60\;{\mu}M$ of $H_2O_2$, respectively. The $H_2O_2$ was on cultured human skin melanoma cells because MCV of $H_2O_2$ was lower than $100\;{\mu}M$. In the antioxidant effect of CRP extract, CRP extract increased cell viability DPPH-radical scavenging activity and SOD-like activity. From these results, it is suggested that $H_2O_2$ was very toxic on cultured human skin melanoma cells. And also, CRP extract has the antioxidant effect on $H_2O_2$-induced cytotoxicity.

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

• Journal of Periodontal and Implant Science
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• v.35 no.4
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• pp.1097-1108
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• 2005

The present study was to determine the influence of micro-macro biphasic calcium phosphate(MBCP) on proliferation and differentiation of human marrow-derived mesenchymal stem cells. Primary stem cells were cultured from bone marrow and 3-4 passaged cells were used. This study tested the proliferative effects by cell counting. Collagen sythensis, alkaline phosphatase activity, expression of osteocalcin and bone sialoprotein by Western blot analysis were evaluated. The cellular proliferation of ASC was not influenced by MBCP. Collagen synthesis of ASC cultured on MBCP significantly increased at 5th and 7th days(p<0.05). The ALP activity in ASC cultured on MBCP significantly increased at 5th and 7th days(p<0.05). The expression of OC and BSP incresaed in ASC cultured on MBCP. These results suggest that MBCP may stimulates the osteoblastic activity of ASC.

• YAKHAK HOEJI
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• v.42 no.3
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• pp.336-344
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• 1998

Effects of ochratoxin A (OTA) on embryo development were studied in cultured whole embryos from 9.5 day gestation rat for 48 h. OTA (more than $0.5{\mu}g/ml$) induced microcephaly in the cultured rat whole embryos. Protein and DNA content, and DNA synthesis were significantly inhibited by OTA. We next examined whether the microcephaly seen in cultured whole embryo partially results from inhibition of differentiation of embryonic midbrain cells. Embryonic midbrain cells were extracted from 12 day gestation rat embryos, and cultured for 96 hr. OTA ibhibited cell differentiation about 50% over control. We also tested whether OTA-induced embryotoxicity would be associated with oxidative damages. We measured the ${\gamma}$-glutamyltranspeptidase (${\gamma}$-GT) and glutathione peroxidase (GPX) activities, and glutathione (GSH) content in both cultured whole embryos and embryonic midbrain cells. OTA decreased GSH content, whereas slightly increased ${\gamma}$-GT activity, but GPX activity was not significantly changed. These results show that OTA caused the microcephaly and its effect may be partially due to the inhibition of cell differentiation of embryonic midbrain cells, but the role of oxidative damages is not clear in embryotoxicity.

• Korean Journal of Animal Reproduction
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• v.16 no.4
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• pp.311-316
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• 1993

Bovine follicular oocytes were matured, fertilized and cultured in vitro. Oocytes with or without cumulus cells were transferred into TC-I99 medium supplemented with 10% FCS 8 h after insemination, and embryos cleaved to 2- to 8-cell stage 56 h after insemination were cultured further for 5 days. The proportions of embryos developed to morular and blastocyst stages were significantly higher (P<0.0l) in those cultured with(31%) than without(15%) cumulus cells. When the embryos were cultured with cumulus cells in the medium with different protein sources, the highest proportion(59%) of embryos developed to morular or blastocyst stage was obtained in medium with BSA(P<0. O01). Higher proportions of embryos were degenerated during culture with FCS(15%) or CS(14%) compared with BSA(3%). The present results indicate the early development of in-vitro fertilized bovine embryos can be maintained efficiently by BSA when they were co-cultured with cumulus cells.

• Journal of Periodontal and Implant Science
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• v.36 no.3
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• pp.757-765
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• 2006

Human periodontal ligament fibroblasts (hPDLF) are very important for curing the periodontal tissue because they can be differentiated into various cells. A tissue engineering approach using a cell-scaffold is essential for comprehending today's periodontal tissue regeneration procedure. This study examined the possibility of using an acellular dermal matrix as a scaffold for human periodontalligament fibroblast (hPDLF). The hPDLF was isolated from the middle third of the root of periodontally healthy teeth extracted for orthodontic reasons. The cells were cultured in a medium containing Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at $37^{\circ}C$ in humidified air with 5% $CO_2$. The acellular dermal matrix(ADM) was provided by the US tissue banks(USA). Second passage cells were used in this study. The hPDLF cells were cultured with the acellular dermal matrix for 2 days, and the dermal matrix cultured by the hPDLF was transferred to a new petri dish and used as the experimental group. The control group was cultured without the acellular dermal matrix, The control and experimental cells were cultured for six weeks. The hPDLF cultured on the acellular dermal matrix was observed by Transmission Electron microscopy (TEM). Electron micrography shows that the hPDLF was proliferated on the acellular dermal matrix. This study suggests that the acellular dermal matrix can be used as a scaffold for hPDLF.

• Journal of Plant Biology
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• v.34 no.1
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• pp.31-35
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• 1991

Induction of symbiosis between N. muscorum and cultured tobacco cells associately cultured on nitrogen-free medium and effects of polyamines on culture condition were carried out. Analysis of amino acid composition in associate cultures showed increase in total amino acid amounts than single culture of cultured tobacco cells and methionine was markly increased in associative culture on N-free medium treated with 1 mM spermine. These results indicated that the composition of amino acids increased effectively in associative cultures by nitrogen fixation of N. muscorum.scorum.

• Korean journal of food and cookery science
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• v.20 no.3
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• pp.287-291
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• 2004

The growth inhibitory effect of germanium, or soybean sprouts cultured with germanium, on cancer cells was determined in the cultured gastric cancer cell line, AGS. The growth of AGS was significantly inhibited by the addition of 0.01-1％ organic germanium (Ge-132) and germanium stone powder in MTT cytotoxicity assays. The juice from germanium treated soybean sprouts (GTS) inhibited the growth of AGS gastric cancer cells by 78-88％ at concentrations of 2.5 or 5${\mu}\ell$. The juice from Seomoktae GTS revealed an especially higher growth inhibitory effect than that from the control soybean sprouts (germanium non-treated soybean sprouts, GNTS) in AGS. The results suggest that soybean sprouts cultured with germanium may exert an anticancer effect against gastric cancer cells.