• KSBB Journal
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• v.7 no.2
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• pp.132-138
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• 1992

Sprial adaptation technique of conditioned media has been applied to cultivate human cell line which can not survive in a serum free mdium without adding any growth factors in basal medium Doubling time and scu-PA production from serum free adapted cells were 5 days and 890 (IU/mL), respectively in a T-flask, whose values were not much lower than the productivity of 1100(IU/mL) from 5% serum containing medium. It was required to use conditioned media for attaching cells on microcarriers when cells were inoculated into a spinner vessel. Then, cells could continuously grow in serum free medium with having specific growth rate of 0.106 (1/day) and specific scu-PA production rate of $1.58{\times}10_{-5}$(IU/cell/day) in batch cultivation.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.1
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• pp.77-81
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• 1994

To improve in vitro embryonic cell development, this study was desigend to culture in vitro fertilized early embryos of mouse in two different systems; conditioned medium alone and ampullary cells co-culture. Thirty two of 83 embryos(38.6%) were blocked in the 2 cell stage by co-culture, as compared to forty of 42 embryos(95.2%) in control group for 24hours culture. And all the embryonic cells cultured for conditioned medium alone were blocked for 48 hours culture. Twenty seven of 46 embryos (58.7 %) which overcome culture block in 2 cell stage by cocultured were developed morular and expanded blastocyst, and ninteen of 46 embryos(26.1 %) underwent hatching for 96 hours culture. The cellular fragmented rates for embryo were 26.2% in medium alone; 10 fragmented blastomere were graded mild status and 1 fragmented blastomere in severe status. On the other hand, the fragmented rate for 48 hours co-cultured were 15.7%03/83); 8 fragmented embryos were graded mild status, moderate status in 3 fragmented embryos and severe in 2 fragmented embryos respectively. In conclusion, the co-culture of embryos with ampullary cells is good to improve quality of embryos and overcome of culture block as well as development of cell cleavage.

• The Journal of Korean Medicine
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• v.26 no.1 s.61
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• pp.1-10
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• 2005

Objective: We examined the antimetastatic effect of Curcuma zedoaria Roscoe (CZ) on pulmonary metastasis of B 16 cells. Methods: For 6 weeks, Zedoariae Rhizoma made from dried CZ were dissolved in distilled water and administered to mice 2 weeks before they were injected with B]6 melanoma cells. Mice were given CZ at doses of 250 and 500 mg/kg, and were compared for lung weight, survival days, and NO production. Results: Intake of CZ throughout the experiment extended the average survival time. Intake after B16 cell injection slightly prolonged survival time, but intake before B]6 cell injection did not influence life span. We examined the effect of CZ on macrophage function by measuring NO production. After the macrophages were given CZ for 6 weeks, the amount of NO generated by the macrophages stimulated with LPS in culture medium increased. NO generated by the macrophages also served as a cytotoxic factor against B16 melanoma cells. B16 melanoma-conditioned medium reduced NO production by macrophages. However, CZ treatment reversed the reduction in NO production by the conditioned medium significantly. Conclusion : These findings may suggest that macrophage function-modulating activity by CZ appears to underlie its antimetastatic activity, which leads to a decrease in the number of lung metastatic surface nodules and the extension of life span.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.10
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• pp.1403-1411
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• 2002

The aims of this study were, firstly, to analyze the biochemical compositions of serum and follicular fluid (FF) from prepubertal gilts after PMSG (1,000 IU) treatment. The concentrations of total proteins, lipids, cholesterol, glucose and sex hormones (progesterone, $P_4$; estradiol-$17{\beta}$, $E_2$; testosterone, T) were measured. Secondary, the effects of porcine FF (pFF) addition (40% and 100%) in IVM media and different culture conditions [Exp. 1: mBMOC-2+20% porcine serum (PS), fresh IVM medium, filtered IVMconditioned medium, or rabbit oviducts; Exp. 2: mBMOC-2+20%PS or stepwise medium replacement procedures (SMRP) cocultured with or without cumulus cells] on the in vitro development (IVD) of porcine oocytes were also examined. Results showed that no significant differences were found in total protein levels between serum and pFF from different sizes (large, >7 mm; medium, ~5-7 mm; small, <3-5 mm) of follicles (75-85 and 49-90 mg/dl; p>0.05). Total lipid concentrations remained constant in serum (395-472 mg/dl), and reduced significantly in the pFF from large follicles (287 mg/dl) at 132 h after PMSG treatment when compared to those at other time points (441-480 mg/dl). Basal cholesterol levels in serum and pFF at 12 h were similar (153-161 mg/dl), but increased at 36 h (186-197 mg/dl). Basal P4 and E2 levels in serum (0.1 ng/ml and 5.5 pg/ml) were low, but increased from 0.34 ng/ml and 12.13 pg/ml at 24 h to 0.81 ng/ml and 61.70 pg/ml at 98 h, respectively, after PMSG treatment (p<0.05). P4 levels increased linearly in pFF from large follicles during 12 through 132 h (138-1,288 ng/ml). A similar increase was also observed in $E_2$ levels (22-730 pg/ml) before 60 h post PMSG treatment, and then dropped afterwards (730-121 pg/ml). The development of the oocytes fertilized in 40% pFF-medium was greater than that in 100% pFF-medium group without gonaodtropin addition (31% vs 10%, p<0.05). However, both were lower than those in mBMOC-2+20%PS and in rabbit oviducts (p<0.05). When cocultured with cumulus cell monolayers, a greater cleavage rate was observed in the group cultured in filtered IVM-conditioned medium than the SMRP group (36% vs 18%, p<0.05). A similar phenomenon was also observed in the culture without cumulus cell monolayers (33% vs 19%, p<0.05). It is concluded that neither the fresh IVM nor filtered IVM-conditioned medium has positive effect on the IVD of oocytes. Coculture with cumulus cell monolayers and the SMRP were not beneficial to the development of IVF pig oocytes.

• Journal of Life Science
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• v.34 no.3
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• pp.170-178
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• 2024

Adipose-derived stem cells (ADSCs) are capable of differentiation into multiple lineages of cells, which has attracted attention for clinical therapy. However, ADSCs have poor proliferation capacity and a short life span in culture, which is an impediment in the application to clinical use. Previously, to overcome growth disadvantages, we had established an immortalized ADSC line (ADSC-T) by introducing the SV40 T antigen coding gene into primary human ADSC. In the present study, we evaluated the differentiation potential of this cell line and assessed the anti-inflammatory effect of its conditioned medium (CM). ADSC-T appeared to maintain the differentiation potential into adipocyte and chondrocyte. The CM of ADSC-T suppressed the NF-κB activity and its target gene expression of COX-2 and iNOS. Furthermore, the phosphorylations of MAPKs, including ERK, JNK and p38, were suppressed by the ADSC-T CM. The expressions of pro-inflammatory cytokines such as TGF-β, TNF-α, IL-6, and IL-13 were also suppressed by the CM of ADSC-T. In the Nc/Nga atopic model mice, the CM showed therapeutic effect on DNCB-induced atopic dermatitis. These results indicate that the immortalized ADSC-T maintains the beneficial properties of primary ADSC and could be a versatile cell source for not only research into ADSC but also for production of CM suitable for clinical application.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.251-251
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• 2004

Plasminogen activators (PA)는 다수의 세포 형태에서 분비되는 것으로 알려진 serine protease이다. PA는 섬유소 용해, 배란, 유선 퇴화, 착상 및 수정 등 다양한 생리적인 과정에 관여한다. 본 연구는 난포액이 돼지 난자의 체외성숙에 미치는 영향을 검토하기 위하여, 다양한 조건하에서의 돼지 난자의 성숙과 난구세포-난자 복합체(Cumulus-Oocyte complexes: COCs) 또는 conditioned medium 내의 PA 활성을 검토하였다. 직경 2∼6m 난포로부터 COCs를 회수하여 일부는 난구세포를 제거하였다. (중략)

• Biomolecules & Therapeutics
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• v.4 no.1
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• pp.46-50
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• 1996

Inhibition of the growth of human cancer cells by proteins secreted from 373-L1 cells was investigated in the present study. The growth of human cancer cells was inhibited by co-culture with 373-L1 cells under 10% FBS and DME, DME, GIT and serumless medium, respectively. The conditioned medium of cultured 373-L1 cells under serumless medium was concentrated 100-fold through an ultrafiltration cell with a 10,000 molecular weight cutoff at 4$^{\circ}C$ under positive pressure using nitrogen(373-L1 EM). 373-L1 EM inhibited the growth of HeLa, Hep G 2, KHOS-Np, A43l and MCF-7 cells. 3T3-L1 EM was purified with FPLC, DEAE-ion exchange chromatography and phenyl-sepharose chromatography. The major protein of 373-L1 EM has a molecular weight of 66,000-68,000 in SDS-PAGE analysis. The results suggest that the inhibitory activity of 373-L1 EM appears to be due to some protein(m.w.66,000-68,000) secreted by 373-L1 cells.

• Animal cells and systems
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• v.2 no.2
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• pp.239-242
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• 1998

Arabidopsis trp1 mutant plants, deficient in phosphoribosyI anthranilate transferase (PAT) activity, accumulate anthranilate compounds, which render them blue fluorescence. The visible phenotype of trp1 makes the PAT gene an excellent reporter gene in the mutant. In order to develop a system for the homologous recombination using the phenotypic characteristic of trp1-100, we established optimum conditions for the isolation and regenera tion of protoplast from auxin-conditioned, trp1-100 root cultures. Trvptophan had to be supplemented in the germination medium for the efficient cell division and subsequent plant regeneration. When 10 uM tryptophan was added to the germination medium, we obtained the highest yield of protoplasts ($3{\times}10^6 cells/g$) and the best viability (92%). Thirty percent of root protoplast derived from meristematic cells underwent cell division within 5 days in callus-induction medium. Regenerated rosette leaves (2-3 mm) were transferred to rooting medium and finally acclimated to the soil for flowering.

• Korean Journal of Plant Tissue Culture
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• v.24 no.3
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• pp.183-188
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• 1997

Callus from scale segments of Lilium longiflorum 'Gelia' was effectively induced and maintained from unorganized tissue on the semi-solid medium by 0.42% Bacto agar with MS basal salts and vitamins of SH medium supplemented with 0.5 mg/L 2, 4-D, 1.0 mg/L NAA, 0.3 mg/L BA, and 3% sucrose. More than 5% of high sucrose level had inhibiting effect on regeneration capacity of formed callus and decreased callus growth. Various combinations of nitrogen did not effective to proliferate the ELC (Embryogenic-like callus), but friability of callus was increased in the medium containing only nitrate as nitrogen source. 5 mL conditioned medium into 30 mL fresh medium was good for cell growth. However friable cell aggregates during suspension culture had to form hard callus which hindered to establish suspention culture system. Addition of 2 g/L casein hydrolysate increased callus growth and friability of the hard callus. As a result of anatomical observation of callus, organogenesis such as shoots, roots and bulblets was independently induced from callus tissue. Somatic embryogenesis from callus tissue could be observed with low frequency.

• Journal of Nutrition and Health
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• v.36 no.3
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• pp.270-279
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• 2003

Conjugated linoleic acid (CLA) is a group of positional and geometric isomers of linoleic acid (LA) and exhibits anticarcinogenic activity in a variety of animal models. We have previously observed that CLA inhibited the growth of Caco-2 cells, a human colon adenocarcinoma cell line. The present study was performed to determine whether the growth inhibitory effect of CLA is related to change in secretion of IGF- II and/or IGF-binding proteins (IGFBPs) that have been shown to regulate Caco-2 cell proliferation by an autocrine mechanism. Cells were incubated in serum-free medium with various concentrations of CLA or linoleic acid (LA). Immunoblot analysis of 24-hours, serum-free, conditioned medium using a monoclonal anti-IGF-IIantibody revealed that Caco-2 cells secreted both mature 6,500 Mr and higher Mr forms of pro IGF-II. The levels of pro IGF-II and mature IGF-IIwere decreased by 43 $\pm$ 2% and 53 $\pm$ 6%, respectively by treatment with 50 $\mu$ M CLA. LA slightly increased pro IGF- II levels. Results from Northern blot analysis showed that CLA decreased IGF-II mRNA levels at 50 $\mu$ M concentration suggesting that CLA regulation of IGF-II protein expression occurs partly at the transcriptional level. Ligand blot analysis of conditioned media using 1251-IGF-II revealed that CLA slightly decreased IGFBP-2 levels and increased IGFBP-4 levels. We confirmed our previous results that CLA inhibited cell growth in a dose-dependent manner but LA slightly increased cell growth. Exogenous IGF-II mitigated the growth inhibitory effect of CLA. These results indicate that the growth inhibitory effect of CLA may be at least in part mediated by decreasing IGF-II and IGFBP-2 secretion and increasing IGFBP-4 secretion in Caco-2 cells.