• Korean Journal of Animal Reproduction
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• v.27 no.1
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• pp.53-59
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• 2003

For somatic cell nuclear transfer in Hanwoo, fetal fibroblast cell lines were established from 35, 50, 70 and 90-day fetuses of Korean native cattle. The sex of these fetal fibroblast cells were analyzed by PCR using Y-specific primers and confirmed that two cell lines were female and the other two cell lines were male. Karyotyping of these cell lines indicates that the chromosome numbers of fetal fibroblast cells were not affected by passage number and more than 80% of fetal fibroblast cells have normal chromosome number. To evaluate Go stage in cell cycle of fetal fibroblast cells, Western blotting was performed to detect the expression level of PCNA which is known to be expressed in all cell cycle stages except G$_{0}$ stage. Following serum starvation or confluent culture for 7 days, fetal fibroblast cells were effectively reached to G$_{0}$ stage. The cell cycle was resumed after culture of these Go stage-fetal fibroblast cells with normal medium. These results indicates that fetal fibroblast cells originated from Hanwoo were successfully isolated and culture system and induction of cell cycle of these cells were established for somatic cell nuclear transfer in Hanwoo.woo.

• Journal of Periodontal and Implant Science
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• v.26 no.2
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• pp.414-428
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• 1996

The use of basic fibroblast growth factor which function as potent biologic mediators regulating numerous activities of wound healing has been suggested for the promotion of periodontal regeneration. The mitogenic effects of basic fibroblast growth factor on human periodontal ligament cells and human gingival fibroblasts were evaluated by determining the incorporation of 5-Bromo-2'deoxy-uridine into DNA of the cells in a dose -dependent manner. The cells which were prepared were the primary cultured gingival fibroblasts and periodontal ligament cells from human the fourth or sixth subpassages were used in the experiments. The cells which were seeded DMEM contain 10% FBS. The added concentrations of basic fibroblast growth factor were 0.1, 1, 10, 50, $l00{\eta}g/ml$ and basic fibroblast growth factor were added to the quiescent cells for 24 hours, 48 hours and 72 hours. They were labeled with $10{\mu}l/200{\mu}l$ 5Bromo-2'-deoxy-uridine for the last 6 hours of each culture. The results of the five determinants were presented as mean and S.D.. The results were as follows. : The DNA synthetic activity of human gingival fibroblasts was increased dose dependently by basic fibroblast growth factor at 24 hours, 48 hours and 72 hours. The similar mitogenic effects were at the 24 and 48 hours of basic fibroblast growth factor, but the DNA synthetic activity of human gingival fibroblasts generally decreased at 72 hours. The DNA synthetic activity of human periodontal ligament cells was increased dose dependently to $50{\eta}g/ml$ by basic fibroblast growth factor at 24, 48 and 72 hours, but the DNA synthetic activity decreased at $l00{\eta}g/ml$ of each hour. Generally the maximum mitogenic effects were at the 48 hours application of basic fibroblast growth factor. The DNA synthetic activity of human periodontal ligament cells generally decreased lower at 72 hours than at 24, 48 hours the application of basic fibroblast growth factor. In the comparison of DNA synthetic activity between human gingival fibroblasts and human periodontal ligament cells, human periodontal ligament cells had slightly higher proliferation activity than human gingival fibroblasts for a longer time at the high dosage of the basic fibroblast growth factor.In conclusion, basic fibroblast growth factor have important roles in the stimulation of DNA synthesis in human periodontal ligament cells and human gingival fibroblasts, and thus may be useful for clinical applications in periodontal regenerative procedures.

• Journal of Periodontal and Implant Science
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• v.36 no.3
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• pp.783-796
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• 2006

The periodontium is a topographically complex organ consisting of epithelial tissue, soft and mineralized tissues. Structures comprising the periodontium include the gingiva, periodontal ligament (PDL) , cementum and the alveolar bone. The molecular mechanism of differentiation in PDL fibroblast cells remain unclear. Amino acid transporters play an important role in supplying nutrition to normal and cancer cells and for cell proliferation. Amino acid transport system L is a major nutrient transport system responsible for the Na+-independent transport of neutral amino acids including several essential amino acids. The system L is divided into two major subgroups, the L-type amino acid transporter 1 (LAT1) and the L-type amino acid transporter 2 (LAT2). In this study, the expression pattern of amino acid transport system L was, therefore, investigated in the differentiation of PDL fibroblast cells. To determine the expression level of amino acid transport system L participating in intracellular transport of amino acids in the differentiation of PDL fibroblast cells, it was examined by RT-PCR, observation of cell morphology, Alizaline red-S staining and uptake analysis after inducing experimental differentiation in PDL fibroblast cells isolated from mouse molar teeth. The results are as follows. 1. The LAT1 mRNA was expressed in the early stage of PDL fibroblast cell differentiation. This expression level was gradually reduced by differentiation- inducing time and it was not observed after the late stage. 2. The expression level of LAT2 mRNA was increased in time-dependent manner during differentiation induction of PDL fibroblast cells. 3. There was no changes in. the expression level of 4F2hc mRNA, the cofactor of LAT1 and LAT2, during differentiation of PDL fibroblast cells. 4. The expression level of ALP mRNA was gradually increased and the expression level of Col I mRNA was decreased during differentiation of PDL fibroblast cells. 5. The L-leucine transport was reduced by time from the early stage to the late stage in PDL fibroblast cell differentiation. As the results, it is considered that among neutral ammo acid transport system L in differentiation of PDL fibroblast cells, the LATl has a key role in cell proliferation in the early stage of cell differentiation and the LAT2 has an important role in the late stage of cell differentiation for providing cells with neutral amino acids including several essential amino acids.

• Biotechnology and Bioprocess Engineering:BBE
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• v.3 no.2
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• pp.51-54
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• 1998

It has been proved that co-cultivation of human neroblastoma cells and human fibroblast cells can enhance nerve cell growth and the production of BDNF in perfusion cultivation. In batch co-cultivation, maximum cell density was increased up to 1.76${\times}$106 viable cells/mL from 9${\times}$105 viable cells/mL of only neuroblastoma cell culture. The growth of neuroblastoma cells was greatly improved by culturing both nerve and fibroblast cells in a perfusion process, maintaining 1.5${\times}$106 viable cells/mL, which was much higher than that form fed-batch cultivation. The nerve cell growth was greatly enhance in both fed-batch and perfusion cultivations while the growth of fibroblast cells was not. It strongly implies that the factors secreted from human fibrobast cells and/or the environments of co-culture system can enhance both cell growth and BDNF secretion. Specific BDNF production rate was not enhanced in co-cultures; however, the production period was increased as the cell growth was lengthened in the co-culture case. Competitive growth between nerve cells and fibroblast cells was not observed in all cases, showing no changes of fibroblast cell growth and only enhancement of the neuroblastoma cell growth and overall BDNF production. It was also found that the perfusion cultivation was the most appropriate process for cultivating two cell lines simultaneously in a bioreactor.

• Journal of Microbiology and Biotechnology
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• v.14 no.1
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• pp.193-196
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• 2004

The peptide Arg-Gly-Asp (RGD) was immobilized through their amino terminus to ends of a sugar bearing copolymer, producing a functional hybrid copolymer. Poly(N-p-vinylbenzyl-D-maltonamide-co-6-(p-vinylbenzamido)-hexanoic acid-g-GRGDS) [p(VMA-co-VBGRGDS)] promoted the attachment and growth of NIH fibroblast cells. The interaction between fibroblast cells and p(VMA-co- VBGRGDS) copolymer resulted in effective cell attachment, proliferation, and morphological changes by introduction of a GRGDS sequence. Moreover, when pretreated with soluble RGD monomer, attachment of fibroblast cells was suppressed approximately 50% from that of the p(VMA-co-VBGRGDS) surface.

• Archives of Plastic Surgery
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• v.37 no.1
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• pp.7-11
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• 2010

Purpose: Vascular endothelial growth factor (VEGF) is known as a growth factor of endothelium and fibroblast. The purpose is to know the VEGF effects on fibroblast proliferation and fibroblast's notch receptor expression. Methods: CCD-986sk fibroblast was purchased from the Korean Cell Bank and was used in XTT assay for proliferation and wound healing assay for migration. Immunofluorescent (IF) staining and western blotting were used in testing notch expression of fibroblast. Semiquantitative RT-PCR was used in checking notch 1 mRNA production by fibroblast. Student-t test was used for analyzing results. Results: Cell proliferation assay using XTT showed significant higher proliferation in VEGF treated fibroblast, $2.324{\pm}0.0026$ vs. $2.463{\pm}0.017$ (p=0.002). Wound healing assay showed longer migration in VEGF treated fibroblast (p=0.062). The fluorescence was brighter in VEGF treated cells of notch 1 IF staining. Notch 1 expressions and mRNA productions increased more in VEGF treated cells. Conclusion: VEGF stimulates fibroblast to proliferate, migrate and to express Notch 1 simultaneously. Notch receptor could be related to VEGF mediated wound healing.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.6
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• pp.998-1005
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• 1997

Kimchi showed protective effect from oxidative damage generated by hydrogen peroxide and paraquat. To investigate the major components of kimchi which reduce the cytotoxicity against reactive oxygen species, keratinocyte(A431, epidermoid carcinoma, human) and fibroblast(CCD-986SK, normal control, human) were cultured under oxidative stress condition provoked by paraquat, a superoxide anion generator, and hydrogen peroxide in the absence or presence of kimchi ingredients. Most keratinocyte and fibroblast cells were killed by hydrogen peroxide and paraquat over 1mM concentration, but kimchi ingredients showed protective effects from oxidative damage generated by hydrogen peroxide and onion, among those, garlic showed the most remarkable preventive effect. Most of kimchi ingredients showed protective effect against paraquat, especially leek notably increased cell survival. For fibroblast cells, ginger had the preventive effect against paraquat, especially leek notably increased cell survival. For fibroblast cells, ginger had the preventive effect from cell killing by high dose of hydrogen peroxide, but most ingredients were not effective against paraquat.

• Journal of The Korean Radiological Technologist Association
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• v.25 no.1
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• pp.317-323
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• 1999

The response of endothelial cells to ionizing radiation is thought to be an important factor in the overall response of normal tissue. It has been reported that basic fibroblast growth factor (bFGF), a potent mitogen for endothelial cells, protects endoth

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.64-64
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• 2001

To facilitate the widespread application of somatic cell cloning, improvements in blastocyst production efficiency and subsequent fetal viability are required. Area where technical improvements are needed include donor cell treatments, starvation and passage numbers. This study was carried out to investigate the effect of serum-starvation and passage on the development of ear skin fibroblast cells cloned embryos. A skin biopsy was obtained from the ear of a 2-year-old Korean Hanwoo female. The cells were cultured in 10% FBS＋DMEM up to 2-3 months(up to 10 passages) and then used. In Experiment 1, the Korean bovine Ear Skin Fibroblast cells (KbESF) were either serum starved (culture in 0.05% FBS＋DMEM) or serum fed (10% FBS＋DMEM) for 4-7 days Prior to NT In Experiment 2, the KbESF cells used for nuclear transfer in these experiments were from passages 2 to 10. The development of 208 nuclear transfer (NT) embryos reconstructed from either serum starved or serum fed ear skin fibroblast was assessed. NT embryos reconstructed from serum starved and serum fed cells showed the same developmental rate (cleavage 80.16 vs. 85.37%; blastocyst 20.63 vs. 19,51%). The development of 590 nuclear transfer (NT) embryos reconstructed from passage 2 to 10 was assessed. We observed the same developmental rates for embryos derived from later Passages as compared with those embryos from early passages(blastocyst from 16.69 to 27.91%, average 20.17%). There was no significant difference between serum-fed and serum-starved donor cells. We observed no difference in developmental rates for embryos derived from 2 to 10 passages. These data show that prolonged culture and serum starvation does not affects the cloning competence of adult somatic cells.

• Journal of Veterinary Clinics
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• v.23 no.2
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• pp.153-157
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• 2006

The purpose of this study was to determine the effects of mitomycin C on conjunctival fibroblast proliferation and apoptosis. Rabbit conjunctival fibroblast mono layers were treated with 48 hours application of mitomycin-C. The viability of cells was evauated by MTT assay. To examine the effect of mitomycin-C on the cell proliferation, immunocytochemistry for BrdU staning was performed. Then, we investigated whether mitomycin-C induced cell's apoptosis by TUNEL staining. As a result of MTT assay, the viability of cells was gradually inhibited in a dose dependent manner by mitomycin-C. The BrdU staining showed that mitomycin-C significantly suppressed the proliferation of conjunctival fibroblast at concentrations of 0.02% or more. When TUNEL assays were performed, the number of apoptotic cells increased 5.6-, 18.5-, and 33.8-fold compared with the control at 0.01, 0.02, and 0.04%, respectively, of mitomycin-C at 48 hours of exposure. Therefore, mitomycin-C may be useful as a regulator in the treatment of corneal diseases that manifest with scar formation and tumor of fibroblast expansion.