• IMMUNE NETWORK
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• v.5 no.1
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• pp.36-44
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• 2005

Background: The anti-tumor therapeutic effect of autologous tumor cell lysate pulseddendritic cells (DCs) was studied for non-immunogenic and immune suppressive lung cancer model. To test the possibility as an adjuvant therapy, minimal residual disease model was considered in mouse in vivo experiments. Methods: Syngeneic 3LL lung cancer cells were inoculated intravenously into the C57BL/6 mouse. Autologous tumor cell (3LL) or allogeneic leukemia cell (WEHI-3) lysate pulsed-DCs were injected twice in two weeks. Intraperitoneal DC injection was started one day (MRD model) after tumor cell inoculation. Two weeks after the final DC injection, tumor formation in the lung and the tumor-specific systemic immunity were observed. Tumor-specific lymphocyte proliferation and the IFN-${\gamma}$ secretion were analyzed for the immune monitoring. Therapeutic DCs were cultured from the bone marrow myeloid lineage cells with GM-CSF and IL-4 for 7 days and pulsed with tumor cell lysate for 18 hrs. Results: Compared to the saline treated group, tumor formation was suppressed in 3LL tumor cell lysate pulsed-DC treated group, while 3LL-specific immune stimulation was minimum. WEHI-3-specific immune stimulation occurred in WEHI-3 lysate-pulsed DC treated group, which had no correlation with tumor regression. Conclusion: The data suggest the possible anti-tumor effect of cultured DCs as an adjuvant therapy for minimal residual disease state of lung cancer. The significance of immune modulation in DC therapy including the possible involvement of NK cell as well as antigen-specific cytotoxic T cell activity induction was discussed.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.6
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• pp.1208-1214
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• 1997

This study was designed to know the effect of kimchi on the hematological and immunological parameters in vivo and in vitro, respectively. To study the effects of kimchi on the hematological parameters, rats(S.D., male) were divided into 4 groups and fed diets containing of 3%, 5% and 10% kimchi or kimchi free diet(control) for 6 weeks. The results of CBC(complete blood cell) tests obtained from the bloods of rates were as follows ; In 10% kimchi group, the level of WBC(white blood cells), RBC(red blood cells), Hgb(hemoglobin), Hct(hematocrit) were increased significantly than those of control group(p<0.05). MCV(mean corpuscular volume), one of the red cell indices, was also increased significantly in the animals fed 10% kimchi(p<0.05). RDW(Red cell distritution width) and PCT(plateletcrit) was lowest in 10% kimchi group(p<0.05). To examine the effects of kimchi on immune cell growth in vitro, three types of mouse immune cells-spleen cells, bone marrow cells, thymus cells-were cultured with extracts of salted Chinese cabbage, fresh kimchi and fermented kimchi(for 1 week) for 12 or 20 days. Control was supplemented with PBS(phosphate buffer saline) excluding kimchi extract. The results of spleen cell, bone marrow cell, and thymus cell cultures showed similar tendency: control medium accelerated death of cells, extracts of salted Chinese cabbage reduced the rate of cell death, and extracts of fresh kimchi and fermented kimchi promoted cell growth. From these results, it could be suggested that kimchi possibly has an effect on the hematopoietic ability and increases immune cell development and growth in vivo.

• Reproductive and Developmental Biology
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• v.34 no.1
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• pp.1-6
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• 2010

Mesenchymal stem cells (MSCs) have the multipotent capacity and this potential can be applied for obtaining valuable cell types which can use for cell therapy on various regenerative diseases. However, insufficient availability of cellular source is the major problem in cell therapy field using adult stem cell sources. Recently, human embryonic stem cells (hESCs) have been highlighted to overcome a limitation of adult cellular sources because they retain unlimited proliferation capacity and pluripotency. To use of hESCs in cell therapy, above all, animal pathogen free culture system and purification of a specific target cell population to avoid teratoma formation are required. In this study, we describe the differentiation of a mesenchymal stem cell-like cells population from feeder-free cultured hESCs(hESC-MSCs) using direct induction system. hESC-MSCs revealed characteristics similar to MSCs derived from bone marrow, and undifferentiated cell markers were extremely low in hESC-MSCs in RT-PCR, immunostaining and FACS analyses. Thus, this study proffer a basis of effective generation of specialized human mesenchymal stem cell types which can use for further clinical applications, from xenofree cultured hESCs using direct induction system.

• Journal of Microbiology and Biotechnology
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• v.17 no.7
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• pp.1113-1119
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• 2007

Human bone marrow-derived mesenchymal stem cells (hBMMSCs) must differentiate into osteogenic cells to allow for successful bone regeneration. In this study, we investigated the effects of different combinations of three soluble osteogenic differentiation-inducing factors [L-ascorbic acid (AC), ${\beta}$-glycerophosphate (${\beta}G$), and bone morphogenic protein-2 (BMP-2)] and the presence of a hydroxyapatite (HA) substrate on hBMMSC osteogenic differentiation in vitro. hBMMSCs were cultured in medium containing various combinations of the soluble factors on culture plates with or without HA coating. After 7 days of culture, alkaline phosphatase (ALP) activity, calcium deposition, and osteoprotegerin (OPG) and osteopontin (OPN) expression were measured. The effects of individual and combined factors were evaluated using a factorial analysis method. BMP-2 predominantly affected expression of early markers of osteogenic differentiation (ALP and OPG). HA had the highest positive effect on OPN expression and calcium deposition. The interaction between AC, ${\beta}G$, and HA had the second highest positive effect on ALP activity.

• Journal of Periodontal and Implant Science
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• v.28 no.2
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• pp.235-247
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• 1998

Calcium sulfate has a long history of medical use as an implant material. The biocompatibiliry of the material has been clearly established. Bone ingrowth concomitant with resorption occurs rapidly with efficient conduction of bone from particle to particle. Calcium sulfate also has a potential for functioning as a good bamer membrane. The purpose of this study was to compare the biocompatibility of different types of calcium sulfate grafting materials including an expelimental calcium sulfate compound on periodontal ligament cells in vitro as a preliminary test towards the development of a more convenient and useful form of grafting material which could promote regeneration of periodontal tissue. Human periodontal ligament cells were collected from the premolar teeth extracted for orthodontic treatment. cells were cultured in a.MEM culture medium containing 20% FBS, at $37^{\circ}C$ and 100% humidity, in a 5% CO2 incubator. Cells were cultured into 96 well culture plate $1{\times}104$ cells per well with $\alpha$-MEM and incubated for 24 hours. After discarding the medium, those cells were cultured in $\alpha$-MEM contained with 10% FBS alone (control group), in medcal-grade calcium sulfate(MGCS group), in plaster(plaster group), experimental calcium sulfate paste(CS paste group) for 1, 2, 3 day respectively. And then each group was characterized by examining of the cell counting, MTI assay, collagen synthesis. The results \vere as follows. 1. In the analysis of cell proliferation by cell counting, both medical-grdde calcium sulfate group and plaster group showed no stastically significant difference at day 1, 2, 3 accept for plaster group at day 1 compared to control group, but there was stastically significant difference between CS paste group and all other groups at day 1, 2, 3(P<0.05). 2. In the analysis of cytotoxicity by MIT assay, both medical-grade calcium sJlfate group and plaster group showed no stastically significant difference compared to control group at day 1, 2, 3 but there was stastically significant difference between CS paste group and all other groups at day 1, 2, 3(P<0.OS). 3. In the analysis of collagen synthesis by immunoblotting assay, high level was detected for medical-grade calcium sulfate group and plaster group at day 1, 2, 3 compared to CS paste group. On the basis of these results, medical-grade calcium sulfate and plaster was shown to possess biocompatibility whereas the CS paste had unfavourable outcome. This observation shows a need for modification of the materials contained in calcium sulfate paste.

• Proceedings of the Korean Vacuum Society Conference
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• 2011.02a
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• pp.20-20
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• 2011

It has been known that, under certain conditions, application of low-temperature atmospheric-pressure plasmas can enhance proliferation of cells. In this study, conditions for optimal cell proliferation were examined for various cells relevant for orthopaedic applications. Plasmas used in our experiments were generated by dielectric barrier discharge (DBD) with a helium flow (of approximately 3 litter/min) into ambient air at atmospheric pressure by a 10 kV~20 kHz power supply. Such plasmas were directly applied to a medium, in which cells of interest were cultured. The cells examined in this study were human synoviocytes, rat mesenchymal stem cells derived from bone marrow or adipose tissue, a mouse osteoblastic cell line (MC3T3-E1), a mouse embryonic mesenchymal cell line (C3H-10T1/2), human osteosarcoma cells (HOS), a mouse myoblast cell line (C2C12), and rat Schwann cells. Since cell proliferation can be enhanced even if the cells are not directly exposed to plasmas but cultured in a medium that is pre-treated by plasma application, it is surmised that long-life free radicals generated in the medium by plasma application stimulate cell proliferation if their densities are appropriate. The level of free radical generation in the medium was examined by dROMs tests and correlation between cell proliferation and oxidative stress was observed. Other applications of plasma medicine in orthopaedics, such as plasma modification of artificial bones and wound healing effects by direct plasma application for mouse models, will be also discussed. The work has been done in collaboration with Prof. H. Yoshikawa and his group members at the School of Medicine, Osaka University.

• Animal cells and systems
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• v.13 no.4
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• pp.363-370
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• 2009

Since vitamin $D_3$ is an important regulator of osteoblastic differentiation, a presently-established vitamin $D_3$-entrapped calcium phosphate film (VCPF) was evaluated for hard tissue engineering. The entrapped vitamin $D_3$ more rapidly induced bone nodule formation. To characterize the cellular events leading to regulations including faster differentiation, signal transduction pathways were investigated in osteoblastic MG63 cells at a molecular level. Major signaling pathways for MG63 cell proliferation including phosphatidylinositol-3-kinase, extracellular signal-regulated kinase, c-Jun N-terminal kinase and focal adhesion kinase pathways were markedly down-regulated when cells were cultured on calcium phosphate film (CPF) and VCPF. This agreed with our earlier observations of the immediate delay in proliferation of MG63 cells upon culture on CPF and VCPF. On the other hand, the p38 mitogen-activated protein kinase (p38 MAPK) and protein kinase A (PKA) pathways were significantly up-regulated on both CPF and VCPF. CPF alone could simulate differential behaviors of MG63 cells even in the absence of osteogenic stimulation and entrapment of vitamin $D_3$ within CPF further amplified the signal pathways, resulting in continued promotion of MG63 cell differentiation. Interplay of p38 MAPK and PKA signaling pathways likely is a significant event for the promotion of differentiation and mineralization of MG63 cells.

• Molecular & Cellular Toxicology
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• v.4 no.3
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• pp.192-198
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• 2008

A variety of titanium (Ti) and its alloys are used in the clinical procedures of bone regeneration for periodontal and dental implant therapies. This study was performed to determine the effect of different surface dental implant materials on biologic responses of a MG-63 human osteoblast-like cell line. MG-63 cells were cultured on Ti coated with hydroxyapatite (HA), calcium metaphosphate (CMP), anodized (A), which compared with non-coated Ti (control). The appearances of surface of dental implant materials and the morphology of these cells were assessed by scanning electron microscopy (SEM). The gene expression profiles of MG-63 cells cultured on Ti were examined by human cDNA microarray (1,152 elements). The expression of several genes was up- and down-regulated by different surfaces of dental implant materials. Interesting, the genes correlated with cellular adhesion and extra cellular matrix (ECM) formation were enhanced, in accordance surface morphology of the dental implant materials used.

• Journal of Microbiology and Biotechnology
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• v.32 no.1
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• pp.126-140
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• 2022

Stem cells can be applied usefully in basic research and clinical field due to their differentiation and self-renewal capacity. The aim of this study was to establish an effective novel therapeutic cellular source and create its molecular expression profile map to elucidate the possible therapeutic mechanism and signaling pathway. We successfully obtained a mesenchymal stem cell population from human embryonic stem cells (hESCs) cultured on chemically defined feeder-free conditions and treated with connective tissue growth factor (CTGF) and performed the expressive proteomic approach to elucidate the molecular basis. We further selected 12 differentially expressed proteins in CTGF-induced hESC-derived mesenchymal stem cells (C-hESC-MSCs), which were found to be involved in the metabolic process, immune response, cell signaling, and cell proliferation, as compared to bone marrow derived-MSCs(BM-MSCs). Moreover, these up-regulated proteins were potentially related to the Wnt/β-catenin pathway. These results suggest that C-hESC-MSCs are a highly proliferative cell population, which can interact with the Wnt/β-catenin signaling pathway; thus, due to the upregulated cell survival ability or downregulated apoptosis effects of C-hESC-MSCs, these can be used as an unlimited cellular source in the cell therapy field for a higher therapeutic potential. Overall, the study provided valuable insights into the molecular functioning of hESC derivatives as a valuable cellular source.

• Journal of Periodontal and Implant Science
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• v.38 no.4
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• pp.595-602
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• 2008

Purpose: Ti-6Al-4V alloy is widely used as an implant material because of its good biocompatibility and good mechanical property compared with commercial pure titanium. Otherwise, toxicity of aluminum and vanadium in vivo has been reported. Ti-8Ta-3Nb alloy is recently developed in the R&D Center for Ti and Special Alloys and it was reported that this alloy has high mechanical strength, no cytotoxicity and similar biocompatibility to commercial pure titanium, but many studies are needed for its clinical use. In these experiment, we carried out different surface treatment on each Ti-8Ta-3Nb alloy disks, then cultured cell on it and assessed biological response. Materials and Methods: cpTi, Ti-6Al-4V, Ti-8Ta-3Nb alloy disks were prepared and carried out sandblasting and acid etching (SLA) or alkali-heat treatment (AH) on the Ti-8Ta-3Nb alloy disks. We cultured primary rat calvarial cells on each surface and assessed early cell attachment and proliferation by scanning electron microscopy, cell proliferation, alkaline phosphatase activity. Result: The rates of cell proliferation on the cpTi, Ti-8Ta-3Nb AH disks were higher than others (p<0.05) and alkaline phosphatase activity was significantly enhanced on the Ti-STa-8Nb AH disks(p<0.05). Conclusion: Most favorable cell response was shown on the Ti-8Ta-3Nb AH surfaces. It is supposed that alkali-heat treatment of the Ti-8Ta-3Nb alloy could be induced earlier bone healing and osseointegration than smooth surface.