• Journal of Nutrition and Health
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• v.29 no.1
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• pp.97-103
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• 1996

This study was performed to investigate the effects of lipid on cellular senescence, lipid peroxide production, and morphological changes. For this study we used primary skin fibroblasts from neonate rats grown in media various lipid contents. Fibroblasts were cultured until they lost their proliferation potential either in control medium (Dulbecco's modified Eagle's medium supplement with 10% fetal bovine serum) or in media supplemented with various concentrations of lipid-cholesterol rice component from bovine serum. Cumulative population doublings(CPD, as an index of cellular life span), and cellular thiobarbituric acid reactive substances (TBARS, as an index of lipid peroxide) concentrations were measured and morphological changes were observed. CPD were shortened with increasing lipid concentration in media ; 28.12 for cells grown in control medium and 13.42, 11.42, and 6.19 for those grown in 0.1%, 1% and 5% lipid rich components containing media, respectively. Cellular proliferation ratios were those grown in 5% lipid rich components containing media were delayed and they were degenerated soon. TBARS concentrations were increased with increasing concentration of lipid in media. Morphological changes were observed in cells grown in control medium by cellular senescence. Especially lipid droplets were observed in cells grown in 5% lipid rich components containing media. Therefore it seems that lipid contents in media had an effect on cellular proliferation and cellular life span, possibly via lipid peroxide production.

• Journal of Embryo Transfer
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• v.13 no.1
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• pp.43-52
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• 1998

The embryogenesis stimulating activity(ESA) had been shown in co-culture of embryos with bovine oviduct epithelial cell(BOEC) and culture in BOEG-conditioned medium. The present study was undertaken to purify and quantify the embryotropic proteins and to determine the optimum concentration of the embryotropic protein for the proper development of embryos. In BOEC-conditioned medium, five major bands of proteins were detected(66, 53, 40, 32 and 24 kDa) by SDS-PAGE. From these proteins, 288pg of protein that had a 32kDa molecular weight was purified by gel filtration column and perfusion chromatography ion-exchange column. When purified protein was supplemented to the in vitro culture media at various concentrations in protein-free media, 2.5$\mu$g /ml supplement group showed significantly higher rates of embryo development into morula /blastocyst stages than other groups(p<0.05). In conclusion, we purified 32kDa protein from BOEC-conditioned medium and this protein showed optimum embryogenesis stimulating effect at 2.5$\mu$g /ml.

• Archives of Pharmacal Research
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• v.28 no.2
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• pp.220-226
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• 2005

Temperature and medium composition were changed with the aim of increasing growth and erythropoietin (EPO) production in EPO-producing Chinese hamster ovary (CHO) cells. We used the CHO cell line, IBE, and its derivative, CO5, which over-expresses the first two enzymes of the urea cycle, carbamoyl phosphate synthetase I (CPS I) and ornithine transcar-bamoylase (OTC). When supplements were added to the medium at $33\;^{\circ}C$, the growth of IBE and CO5 cells increased by $27\%\;and;26\%$, respectively and the maximum yield of EPO was increased by $40\%$ in both cell lines. The absolute EPO concentration in the CO5 cells was always $55{\sim}60\%$ higher than in the IBE cells. In addition, when the two cell lines were continuously cultured with supplements at $33\;^{\circ}C$ until their growth rates approached those at $37\;^{\circ}C$, the growth rates of both IBE and CO5 cells increased by $54\%$ and their maximum EPO levels increased by up to $73\%\;and\;56\%$, respectively. Therefore, the growth and EPO expression levels of CO5 cells increased 2.2-fold and 2.6-fold, respectively, compared to those of the IBE cells. These results indicate that adaptation to lower temperature as well as medium supplementation could be important for improving cell growth and EPO production.

• Journal of Microbiology and Biotechnology
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• v.16 no.6
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• pp.849-854
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• 2006

The effects of Haematococcus pluvialis extracellular products on microbial growth and bacteriocin production were investigated to improve bacteriocin synthesis during the growth cycle of Lactobacilli. Lactobacillus pentosus KJ-108, L. plantarum KJ-10311, and L. sakei KJ-2008 were cultured in MRS and enriched medium (ERM) with or without supplement of the extracellular products obtained from a late exponential phase culture of Haematococcus pluvialis in modified Bold's basal medium (MBBM). In both MRS and ERM, the extracellular products strongly enhanced the growth as well as the bacteriocin production of all the lactic acid bacteria tested. The enhancing effect was observed in ERM with pH adjusted at 5 and 6. In addition, some difference in growth effects with the extracellular products of H. pluvialis was observed between pH 5 and 6 in ERM, but no effect was observed in the minimal medium. The final biomass and the final concentration of bacteriocin activity were associated with the cell growth that was promoted by the extracellular products of H. pluvialis, and the enhanced cell growth of the three lactic acid bacterial strains induced the increase of the specific bacteriocin production. Therefore, bacteriocin production and activity were influenced by the addition of the extracellular products of H. pluvialis in the culture medium.

• Fisheries and Aquatic Sciences
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• v.18 no.1
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• pp.81-88
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• 2015

We investigated factors affecting primary cultures of Pacific abalone Haliotis discus hannai ovary-dissociated cells to identify general aspects of their early-phase culture. Ninety-seven cell populations derived from 30 individuals were cultured in different media with varying compositions of medium supplements, and initial attachment, subculture, and survival for ${\geq}10$ weeks were assessed according to medium composition and individual. We also examined the time required for subculture and the rate of cell death according to both culturing period and passage number within 10 weeks. A lack of fetal bovine serum (FBS) and hemolymph significantly inhibited the growth of cultured cells, while we detected no significant effect of medium composition on initial cell attachment. Through data reallocation, with the omission of data from cell populations cultured in FBS-free and hemolymph-free media, we showed that growth inhibition was also affected by individual differences among the abalones used. During the culture, we observed four different types of cell morphology. Moreover, considerable time was required for subculture-18.4 and 19.5 days for first and second subcultures, respectively-and cell death did not occur within 30 days or for passage 0. Our results will provide valuable information for developing universal cell culturing guidelines in abalone species and suggest the feasibility of culturing abalone ovary-dissociated cells.

• Korean Journal of Plant Tissue Culture
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• v.28 no.6
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• pp.305-310
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• 2001

The effect of low dose ${\gamma}$-radiation on the callus growth of Lithospermum erythrorhizon S. cultured on different medium and lighting condition was investigated. The 8 Gy irradiation stimulated callus growth on LS medium supplemented with BA 2 mg/L and NAA 2 mg/L, however, the growth of callus was more effective on LS medium supplemented with BA 1 mg/L and NAA 1 mg/L under 16 hrs day light. And on the LS medium containing IAA 0.2 mg/L, 16 Gy irradiation increased the callus growth by supplement with kinetin 2 mg/1 and the effect of kinetin was higher than BA at same concentration. The growth of callus was more vigorous on LS medium than MS medium in general. On M-9 medium, the growth of callus was poor regardless lighting conditions, however, by ${\gamma}$-ray irradiation of 16 Gy or 30 Gy, callus growth rates were increased by 30% or more than 30%, especially, under 16 hrs day light condition.

• Asian-Australasian Journal of Animal Sciences
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• v.9 no.5
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• pp.583-590
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• 1996

The purpose of this study was to evaluate some factors in the bovine embryonic development from one-cell to blastocyst using modified synthetic oviduct fluid medium (mSOFM), after maturation and in vitro fertilization of the oocytes. The embryonic development to the blastocyst stage was assessed at 7-10 days after in vitro fertilization, and the total cells in the blastocysts were counted by staining nuclei with fluorochrome. Some commercial calf sera (CS) and a superovulated cow serum had different effects on the embryonic development to the blastocyst stage (8.6-21.4%), dependent upon their product lots, although the development might not be affected at least by serum progesterone levels. ${\beta}$-Mercaptoethanol (${\beta}$-ME) supplemented into mSOFM was effective to the embryonic development (27.8%), as well as the co-culture system with cumulus cells (19.5%). In a serum- and feeder cell-free culture using mSOFM containing several growth factors and ${\beta}$-ME instead of CS plus co-cultured cumulus cells, bovine serum albumin (BSA, fraction V), but not polyvinyl alcohol (PVA), was highly effective in embryonic development to the blastocyst stage, almost comparable to CS in the serum-contained culture (CS, BSA and PVA; 27.8, 19.5 and 5.7%, respectively). However, fatty acid free BSA rather reduced the number of developed blastocysts, compared with fraction V BSA (7.3 vs 29.4%). In the serum- and feeder cell-free culture, supplement of glucose to the medium (final 2.0 mM) stimulated the cell proliferation of developing embryos 120 hr after in vitro fertilization. These results indicated that a serum-free medium supplemented with ${\beta}$-ME could successfully support the development of bovine one-cell embryos to the blastocyst stage. Moreover, supplement of glucose and fatty acids to the medium might support preferably the development and cell proliferation of embryos.

• Journal of Applied Biological Chemistry
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• v.57 no.1
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• pp.61-63
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• 2014

Aged black garlic (ABG) was extracted with 20% ethanol and water (crude extracts) and fractionated into three categories (>10, 3-10, and <3 kDa). The effect of crude extract supplements on anti-My-10 hybridoma cell growth and IgG1 antibody production was investigated in suspension culture with a chemically defined protein-free medium. We observed that supplementation of ABG to the cell culture medium stimulated anti-My-10 hybridoma cell growth and production of IgG1 antibody, particularly with fractionated ABG of low molecular weight. The stimulation depended upon the concentration and the size of the fractionated ABG. We also found that the growth-promoting activity was not correlated with high antibody production. These results suggest that fractionated ABG is a novel and promising alternative as an animal cell culture supplement.

• Asian Journal of Turfgrass Science
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• v.12 no.4
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• pp.189-194
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• 1998

The effect of proline, glutamine, aspartic acid and their combinations on callus induction and embriogenic callus formation from 3 creeping bentgrass (Agrostis palustris) cv. Regent, Mariner, Cato and 1 colonial bentgrass (Agrostis tenuis) cv. Tiger was estimated in both light and dark condition. The addition of amino acids to the growth medium did not have a significant stimulatory effect on the induction of embryogenic callus, instead, they were inhibitory, particularly at higher concentration (40 mM). But supplement of amino acids at lower concentrations (5 or 10mM) to basal medium was beneficial in inhibiting the formation of hairy outgrowth on the surface of embryogenic callus.

• Journal of Ginseng Research
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• v.6 no.2
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• pp.162-167
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• 1982

Calli and leaflets of ginseng (Panax ginseng C.A. Meyer) were cultured on 1/2MS media supplement. with kinetin, 2 iP, NAA, 2,4-D and IBA to assess their capacity to regenerate embryoids and organs. Root calli produced numerous embryoids and shoots in 1/2MS medium supplemented with 2 mg/l NAA and 2mg/s 2iP, and the combination of 2 iP and NAA was more effective than the combination of kinetin and NAA in induction of embryoid and shoot from root calli. Culture of leaflet in the medium supplemented with IBA resulted in profuse root regeneration.