• Molecules and Cells
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• v.21 no.2
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• pp.206-212
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• 2006

We have established in culture a spontaneously immortalized bovine embryonic fibroblast (BEF) cell line that has lost p53 and $p16^{INK4a}$ functions. MyoD is a muscle-specific regulator capable of inducing myogenesis in a number of cell types. When the BEF cells were transduced with MyoD they differentiated efficiently to desmin-positive myofibers in the presence of 2% horse serum and 1.7 nM insulin. The myogenic differentiation of this cell line was more rapid and obvious than that of C2C12 cells, as judged by morphological changes and expression of various muscle regulatory factors. To confirm that lack of the p53 and $p16^{INK4a}$ pathway does not prevent MyoD-mediated myogenesis, we established a cell line transformed with SV40LT (BEFV) and introduced MyoD into it. In the presence of 2% horse serum and 1.7 nM insulin, the MyoD-transduced BEFV cells differentiated like the MyoD-transduced BEFS cells, and displayed a similar pattern of expression of muscle regulatory proteins. Taken together, our results indicate that MyoD overexpression overcomes the defect in muscle differentiation associated with immortalization and cell transformation caused by the loss of p53 and Rb functions.

• Clinical and Experimental Reproductive Medicine
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• v.32 no.2
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• pp.133-147
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• 2005

Objectives: This study was carried out to establish human embryonic stem cells derived from frozen-thawed embryos using mouse embryonic fibroblasts (mEFs), human fetal skin and muscle fibroblasts as feeder cells, and to identify the characteristic of embryonic stem cells. Methods: When primary mEFs, human fetal skin and muscle fibroblasts were prepared, passaging on 4 days from replating could have effective trypsinization and clear feeder layers. Eight of 23 frozenthawed 4~8 cell stage embryos donated from consenting couples developed to blastocysts. Inner cell mass (ICM) was isolated by immunosurgery. ICM was co-cultured on mEFs, human fetal skin or muscle fibroblasts. The ICM colonies grown on mEFs, human fetal skin or muscle fibroblasts were tested the expression of stage specific embryonic antigen-3, -4 (SSEA-3, -4), octamer binding transcription factor-4 mRNA (Oct-4) and alkaline phosphatase surface marker. Results: We obtained 1 ICM colony from 2 ICM co-cultured on mEFs as feeder cells and did not obtain any ICM colony from 6 ICM clumps co-cultured on human fetal skin or muscle fibroblasts. The colony formed on mEFs could be passaged 30 times every 5 days with sustaining undifferentiated colony appearance. When the colonies cultured on mEFs were grown on human fetal skin or muscle fibroblasts, the colonies could be passaged 15 times every 9 days with sustaining undifferentiated colony appearance. The colonies grown on mEFs and human fetal fibroblasts expressed SSEA-4 and alkaline phosphatase surface markers and positive for the expression of Oct-4 by reverse transcription-polymerase chain reaction (RT-PCR). The produced embryoid body differentiated spontaneously to neural progenitorlike cells, neuron-like cells and beating cardiomyocyte-like cells, and frozen-thawed embryonic stem cells displayed normal 46,XX karyotype. Conclusions: The human embryonic stem cells can be established by using mEFs and human fetal fibroblasts produced in laboratory as feeder cells.

• Korean Journal of Poultry Science
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• v.46 no.3
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• pp.185-191
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• 2019

Nucleoporin 210 (Nup210) is associated with several physiological processes including muscle and neural cell differentiation, autoimmune diseases, and peripheral T cell homeostasis. Chicken Nup210 (chNup210) gene was originally identified as one of the differentially expressed genes (DEGs) in the kidney tissues of chicken. To elucidate the role of Nup210 in metabolic disease of chicken, we studied the molecular characteristics of chNup210 and analyzed its gene expression under the stimulation of Toll-like receptor 3 (TLR3) ligands. The Nup210 genomic DNA and amino acid sequences of various species including fowls, fishes, and mammals were retrieved from the Ensemble database and subjected to bioinformatics analyses. The expression of Nup210 from several chicken tissues was probed through qRT-PCR, and chicken fibroblast DF-1 cell line was used to determine the change in expression of chNup210 after stimulation with TLR3 ligand, polyinosinic-polycytidylic acid (poly (I:C)). The chNup210 gene was highly expressed in chicken lung and spleen tissues. Although highly conserved among the species, chNup210 was evolutionary clustered in the same clade as that of duck compared to other mammals. Furthermore, this study revealed that chNup210 is expressed in TLR3 signaling pathway and provides fundamental information on Nup210 expression in chicken. Future studies that offer insight into the involvement of chNup210 in the chicken innate immune response against viral infection are recommended.

• Molecules and Cells
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• v.24 no.3
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• pp.343-350
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• 2007

Mesenchymal stem/progenitor cells (MPCs) were isolated from porcine umbilical cord blood (UCB) and their morphology, proliferation, cell cycle status, cell-surface antigen profile and expression of hematopoietic cytokines were characterized. Their capacity to differentiate in vitro into osteocytes, adipocytes and chondrocytes was also evaluated. Primary cultures of adherent porcine MPCs (pMPCs) exhibited a typical fibroblast-like morphology with significant renewal capacity and proliferative ability. Subsequent robust cell growth was indicated by the high percentage of quiescent (G0/G1) cells. The cells expressed the mesenchymal surface markers, CD29, CD49b and CD105, but not the hematopoietic markers, CD45 and CD133 and synthesized hematopoietic cytokines. Over 21 days of induction, the cells differentiated into osteocytes adipocytes and chondrocytes. The expression of lineage specific genes was gradually upregulated during osteogenesis, adipogenesis and chondrogenesis. We conclude that porcine umbilical cord blood contains a population of MPCs capable of self-renewal and of differentiating in vitro into three classical mesenchymal lineages.

• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.3
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• pp.425-430
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• 2011

This study evaluated the effect of extraction conditions of yam (Dioscorea aimadoimo) on antioxidant, moisturizing, collagenase activity, proliferation, and migration. Yam has been recognized as a healthy food due to its various biological activities, such as anti-obesity, anti-constipation, anti-mutagenic activities, as well as its ability to decrease blood glucose and cholesterol levels. Electron donating ability of high temperature ethanol extract of Dioscorea aimadoimo (HDA) had shown 70.6% at 400 mg/ml, and low temperature ethanol extract of Dioscorea aimadoimo (LDA) had shown 40% at 400 mg/ml. SOD-like activities of LDA and HDA were 23% and 34% at 400 mg/ml respectively. LDA significantly reduced the activity of collagenase in a dose-dependent manner, which was higher than HDA. The water contents in LDA-treated skin and HDA-treated skin were increased by 45.63% and 38.65% than the placebo cream respectively. The cellular proliferation of human dermal fibroblast neonatal (HDFn) was evaluated by MTT and cell migration assay. Compared to control, the cell proliferation was elevated to 109.7% and 114% by the treatment of LDA and HDA respectively at the concentration of 200 mg/ml. In addition, LDA and HDA were induced cell migration in HDFn. Our study suggests that LDA and HDA should be a very useful cosmetic ingredient, as anti-aging and skin moisturizer.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.1
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• pp.83-87
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• 1999

Since the blastocyst is broken and spreads out on a flat plastic culture dish (two dimensional culture) during in vitro development, it has been difficult to study the implantation process. It also has been difficult to analyse the interactions between endometrial epithelial and stromal cells because of the lack of a long-term in vitro model which can stimulate in vivo characteristics, as these cells eventually fail to proliferate or cease to express differentiated functions. Recently nontransformed cell lines, CUE-P and CUS-V2, derived from rat endometrial epithelium and stroma were reported. In this study, morphology of CUE-P and CUS-V2 was examined and oxytocin gene expression by CUE-P cells was demonstrated by RT-PCR. The CUE-P cells have a cuboidal morphology and CUS-V2 cells resemble fibroblast and exhibit a spindle-like morphology. In RT-PCR, same size of PCR products of oxytocin gene at hypothalamus, uterus and CUE-P cells were demonstrated. These results showed three dimensional culture system could be made by using the new cell lines.

• The Korean Journal of Cytopathology
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• v.4 no.2
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• pp.171-175
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• 1993

Synovial sarcoma us a rare malignant neoplasm of the soft tissue arising in the lower extremity, inguinal area, and upper arm. The majority occurs in patients between the age of 15 and 40 years. The histologic diagnosis is based on the classical biphasic type with the distinct epithelial and spindle cell components. We have recently encountered a case of metastatic synovial sarcoma of the lung diagnosed by fine needle aspiration cytology. A 34-year-old man was admitted because of a palpable mass on the antero-lateral side of the right tibia for 3 years. On admission, a well demarcated metastatic pulmonary nodule, measuring 5 cm in diameter, was also identified in the simple chest X-ray. Resection of the lower leg mass revealed typical histologic features of biphasic synovial sarcoma. Aspiration cytology of the pulmonary nodule revealed numerous clusters of spindle cells admixed with groups of epithelial cells. The epithelial cells had moderate-sized, round to oval shaped, and hyperchromatic nuclei. The cytoplasm was clear, but not distinctive. Interspersed tell elements were fibroblast-like spindle cells having elongated hyperchromatic nuclei.

• Journal of Periodontal and Implant Science
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• v.27 no.3
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• pp.459-468
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• 1997

Healing of periodontal tissues require the migration and proliferation of gingival fibroblasts and periodontal ligament cells. There is many evidences that the some agents like cytokines and polypeptide growth factors are mediate these cellular events in wound healing. Recently someone is interested in herbal drugs on periodontal tissue healing processes. The purpose of this study was to examine the effects of 4 herbal drugs, Carthami Flis, Moutan Redias Cortex, Scirpi Rhisoma, Seed of Carthamus tinctorius L. on human gingival fibroblasts and periodontal ligament cells. Periodontal ligament cells and gingival fibroblasts were primarily cultured from extracted premolar with non-periodontal diseases. The powder from extracted. herbal drugs were prepared with distilled water. Cells were cultured with DMEM at $37^{\circ}C$, 5% $CO_2$, 100% humidity incubator, and treated with each herbal drugs with proper concentration for 1, 2, and 3 days. The cell activity was determined by ELISA reader using MTT assay. There was the most significant elevation in $10^{-3}g/ml$ of almost herbal drugs on cellular activities. The result of this study demonstrated that Carthami Flis, Moutan Radicis Cortex, Scirpi Rhisoma, Seed of Carthamus tinctorius L. appears to have beneficial effect on healing process after periodontal treatment.

• Journal of Korean Foot and Ankle Society
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• v.23 no.2
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• pp.74-77
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• 2019

Fibroma of the tendon sheath (FTS) was initially described in 1936 by Geschickter and Copeland as a benign firmed soft tissue tumor that is rare and less common than another soft tissue tumors, especially giant cell tumors (GCT) of the tendon sheath. The common distinct feature is a slow-growing least painful rare entity arising from the tendon or tendon sheath. FTS is detected mostly in the fingers, hands and wrists but less commonly in the foot. Very few cases of FTS have been described arising from a flexor tendon of the foot. This article describes a 51-year-old patient with FTS that developed in the extensor tendon of the foot, which is the only known FTS to form in this area. Heterogeneous low signal intensity in both the T1- and T2-weighted images was observed in magnetic resonance imaging. The lesion was excised completely by open surgery. Histologically, it showed randomly arranged, fibroblast-like spindle cells in dense fibrous tissue and had insufficient hemosiderin-laden macrophages that are typical for GCT.

• Clinical and Experimental Reproductive Medicine
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• v.35 no.4
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• pp.247-265
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• 2008

Objectives: Many types of liver diseases can damage regenerative potential of mature hepatocytes, hepatic progenitor cells or oval cells. In such cases, a stem cell-based therapy can be an alternative therapeutic option. We examined whether human amnion-derived mesenchymal stem cells (HAM) and human umbilical cord-derived stem cells (HUC) could differentiate into hepatocyte-like cells as therapeutic cells for the liver diseases. Methods: HAM and HUC were isolated from the amnion and umbilical cord of the volunteers after a caesarean section with informed consent. In order to differentiate these cells into hepatocyte-like cells, cells were cultivated in hepatogenic medium using culture plates coated with fibronectin. Effects of hepatocyte growth factor, L-ascorbic acid 2-phosphate, insulin premixture fibroblast growth gactor 4, dimethylsulfoxide, oncostatin M and/or dexamethasone were examined on the hepatic differentiation. After differentiation, the cells were analyzed by RT-PCR, immunocytochemistry, immunoblotting, albumin ELISA, urea assay and periodic acid-schiffs staining. Results: Initial fibroblast-like appearance of HAM and HUC changed to a round shape during culture in the hepatogenic medium. However, in all hepatogenic conditions examined, HUC secreted more amounts of albumin or urea into medium than HAM. Expression of some of hepatocyte-specific genes increased and expression of new genes were observed in HUC following cultivation in hepatogenic medium. Results of immunocytochemistry and immunoblotting analyses demonstrated that HUC secreted albumin into the culture medium. PAS staining further demonstrated that HUC could store glycogen inside of the cells. Conclusions: Both HUC and HAM could differentiate into albumin-secreting, hepatocyte-like cells. Under the same hepatogenic conditions examined, HUC more efficiently differentiated into hepatocyte-like cells compared with the HAM. The results suggest that HUC and HAM could be used as sources of stem cells for the cell-based therapeutics such as in liver diseases.