• Biomaterials and Biomechanics in Bioengineering
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• v.3 no.3
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• pp.141-155
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• 2016

Two-dimensional (2D) cell culture and in vivo cancer model systems have been used to understand cancer biology and develop drug delivery systems for cancer therapy. Although cell culture and in vivo model studies have provided critical contribution about disease mechanism, these models present important problems. 2D tissue culture models lack of three dimensional (3D) structure, while animal models are expensive, time consuming, and inadequate to reflect human tumor biology. Up to the present, scaffolds and 3D matrices have been used for many different clinical applications in regenerative medicine such as heart valves, corneal implants and artificial cartilage. While tissue engineering has focused on clinical applications in regenerative medicine, scaffolds can be used in in vitro tumor models to better understand tumor relapse and metastasis. Because 3D in vitro models can partially mimic the tumor microenvironment as follows. This review focuses on different scaffold production techniques and polymer types for tumor model applications in cancer tissue engineering and reports recent studies about in vitro 3D polymeric tumor models including breast, ewing sarcoma, pancreas, oral, prostate and brain cancers.

• Journal of Microbiology and Biotechnology
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• v.32 no.2
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• pp.238-247
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• 2022

Since the skin covers most surfaces of the body, it is susceptible to damage, which can be fatal depending on the degree of injury to the skin because it defends against external attack and protects internal structures. Various types of artificial skin are being studied for transplantation to repair damaged skin, and recently, the production of replaceable skin using three-dimensional (3D) bioprinting technology has also been investigated. In this study, skin tissue was produced using a 3D bioprinter with human skin cell lines and cells extracted from mouse skin, and the printing conditions were optimized. Gelatin was used as a bioink, and fibrinogen and alginate were used for tissue hardening after printing. Printed skin tissue maintained a survival rate of 90% or more when cultured for 14 days. Culture conditions were established using 8 mM calcium chloride treatment and the skin tissue was exposed to air to optimize epidermal cell differentiation. The skin tissue was cultured for 14 days after differentiation induction by this optimized culture method, and immunofluorescent staining was performed using epidermal cell differentiation markers to investigate whether the epidermal cells had differentiated. After differentiation, loricrin, which is normally found in terminally differentiated epidermal cells, was observed in the cells at the tip of the epidermal layer, and cytokeratin 14 was expressed in the lower cells of the epidermis layer. Collectively, this study may provide optimized conditions for bioprinting and keratinization for three-dimensional skin production.

• Journal of Plant Biology
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• v.27 no.3
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• pp.173-178
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• 1984

Callus culture of Phaseolus vulgaris L. was carried out to examine the ability to grow on cytokinin-free medium. Of the sixteen cultivars of P. vulgaris, eight were classified as completely cytokinin-autonomous phenotype and five were found to be cytokinin-dependent phenotype. Intermediate phenotype was shown in three cultivars. Using cv. Palgong and ca 21 as cytokinin-dependent genotypes, the genotype responses to the cytokinin requirements of callus tissue were studied in detail. The callus tissue of cv. Palgong and ca 21 were never habituated in cytokinin-free medium, regardless tissue origin and cytokinin concentration in previous passages. The result suggests that cytokinin dependency of callus tissue of P. vulgaris cv. Palgong and ca 21 may be due to inactivation of cytokinin biosynthetic pathway.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.2
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• pp.84-91
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• 2000

Mycopasma contamination of tissue culture cells easily evades detection and, thus, represents a continous therat to cell biologists. In case where infected cell can not simply be replaced, attempts have to be made to eradicate mycoplacma from the tissue culture cells. A variety of anti-microbial agents have been shown to be toxic to mycoplasma strains ; however, cell associated mycoplasma are often protected from antibiotics at concentrations shown to be effective in vitro. Antibiotic concentrations high enough to be lethal to cell as sociated mycoplasmas frequently are also detrimentrations to the host cells, while moderately increased antibiotic levels tolerated by the host cells often lead to only temporary growth suppression and/or to the emergence of mycoplasma strains resistanct even to high concentrations of the antibiotis applied. Hare, a genetic approach for the elimination of mycoplasma from tissue culture cells that overcomes thens limitations is described. By expression of a selection marker conferring resistance to an otherwise toxic agent, Acholeplasma laidlawii infected BHK-21 cells used as the model system were enabled to temporarily tolerate antibiotic concentrations high enough to be lethal to cell associated mycopalsma while leaving the host cells unharmed. Upon successful mycoplasma eradicated, cultvation of the cured host cells in the absence of the selective agent yielded revertant cell clones that had regained susceptibillity to the toxic agent. Cressation of the selection marker expression was shown to result from the loss of the selection marker DNA, which is a consequence of the fact that the stable and permanent integration of foreign DNA in eucaryotic cell chrosomes is highly inefficient. Thus, the cells were cured from mycoplasma yet remained biochemically unaltered.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.29 no.4
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• pp.249-256
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• 2003

The lack of sufficient oral mucosa available for intra-oral reconstruction has been dealt with by the use of skin or oral mucosa grafts harvested from donor sites but grafts requires more than one surgical procedures and could cause donor site morbidity. Many investigators have attempted to increase available soft tissue by tissue engineered skin or oral mucosa replacements for clinical applications. But, reconstructed mucosa by several methods have low physical properties such as rolling and contraction. The aims of this study were to develope an in vitro experimental model that maintains an epithelial-mesenchymal interaction by organotypic raft culture, and to characterize biologic properties of three-dimensionally cultured oral mucosa embedded with Polydioxanone mesh by histological and immunohistochemical analysis. The results were as follows; 1. Oral mucosa reconstructed by three-dimensional organotypic culture revealed similar morphologic characteristics to equvalent normal oral mucosa in the point that they show stratification and differentiation. 2. The expression of cytokeratin 10/13 and involucrin in the cultured tissue showed the same pattern with normal oral mucosa suggesting that organotypic co-culture condition is able to induce cellular differentiation. 3. After insertion of polydioxanone mesh, increased tensile strength were observed. These results suggest that three-dimensional organotypic co-culture of the oral mucosa cell lines with the dermal equvalent consisting type I collagen and fibroblasts reproduce the morphologic and immunohistochemical characteristics similar to those in vivo condition. And increased physical properties by use of polydioxanone mesh will helpful for clinical applications.

• Journal of Periodontal and Implant Science
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• v.26 no.1
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• pp.143-160
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• 1996

The purpose of this study was to evaluate the response in aspect of attachment and growth rate of osteoblasts and growth rate of osteoblasts and human gingival fibroblasts to the commercially pure titanium(CP titanium)and titanium alloy(Ti-6AI-4V) that are used widely as implant materials, and to obtain the basic information to ideal implant materials. In the studly, commercially pure titanium in first test group, titanium alloy(Ti-6AI-4V) in second test group, cobalt-chrome-molybdenum alloy(Co-Cr-Mo alloy) in positive control group, and tissue culture polystyrene plate in negative control group were used. The results of this study were as follows. 1. Bone marrow cells cultured on CP titanium and Ti-6Al-4V showed significantly greater attachment and growth rate(p(0.05) compared to Co-Cr-Mo alloy in each time. 2. There were no significant differences(p>0.05) in attachment and growth rate of bone marrow cells cultured on CP titanium and Ti-6AI-4V or tissue culture plate. 3. Most bone marrow cells cultured on CP titanium, Ti-6Al-4V and tissue culture plate were attached well to each substratum in first 2days, and then, grew at higher growth rate. On the other hand, some cells cultured on Co-Cr-Mo alloy failed to attach in first 2 days, and then, attached cells grew at lower growth rate than other groups. 4. Attachment and growth rates of gingival fibroblasts cultured on CP titanium and Ti-6Al-4V showed no significant differences(p>0.05) compared to Co-Cr-Mo alloy in 2 days, but significantly greater increase(p<0.05) in 5 and 9 days. 5. There were no significantly differences(p>0.05) between growth rates on gingival fibroblasts cultured on CP titanium, Ti-6Al-4V and tissue culture plate in 2 and 5days, but a significant lower growth rate(p<0.05) on CP titanium and Ti-6Al-4V versus tissue culture plate. 6. Some gingival fibroblasts cultured on all specimen groups failed to attach, but attached cells grew well, especially on CP titanium, Ti-GAl-4V and tissue culture plate. 7. There were no significant differences(P>0.05) between growth rates of both bone marrow cells and gingival fibroblasts cultured on CP titanium and Ti-6AI-4V. As a result of this study, both commercially pure titanium and Ti-6AI-4V showed excellent biocompatibility and there was no significant difference in the cellular response to the both metals. Bone marrow cells cultured on each substratum showed significantly greater growth rate and responded sensitively to cytotoxic effects of metal surfaces compared to gingival fibroblasts. Considering cell response to the substrate, it was likely that the composition itself of titanium metals have no significant effect on the biocompatibility. Further study need to be done to evaluate the influence of surface characteristics on cellular responses.

• Journal of Microbiology and Biotechnology
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• v.16 no.10
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• pp.1577-1582
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• 2006

Cartilage tissue engineering has emerged as an alternative approach for reconstruction or repair of injured cartilage tissues. In this study, rabbit chondrocytes were cultured in a three-dimensional environment to fabricate a new cartilaginous tissue with the application of tissue engineering strategies based on biodegradable PLGA microspheres. Chondrocytes were seeded on PLGA microspheres and cultured on a rocking platform for 5 weeks. The PLGA microspheres provided more surface area to adhere chondrocytes compared with PLGA sponge scaffolds. The novel system facilitated uniform distribution of the cells on the whole of the PLGA microspheres, thus forming a new cartilaginous construct at 4 weeks of culture. The histological and immunohistochemical analyses verified that the number of chondrocytes and the amount of extracellular matrix components such as proteoglycans and type II collagen were significantly greater on the PLGA microspheres constructs as compared with those on the PLGA sponge scaffolds. Therefore, PLGA microspheres enhanced the function of chondrocytes compared with PLGA sponge scaffolds, and thus might be useful for formation of cartilage tissue in vitro.

• ALGAE
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• v.28 no.1
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• pp.83-92
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• 2013

The descriptions of galls, or tumors, in red algae have been sparse. K$\ddot{u}$tzing (1865) observed possible galls of Bostrychia but only presented a drawing. Intensive culture observations of hundreds of specimens of the genus Bostrychia over many years have revealed that galls appeared in only a small subset of our unialgal cultures of B. kelanensis, Bostrychia moritziana/radicans, B. radicosa, B. simpliciuscula, and B. tenella and continued to be produced intermittently or continuously over many years in some cultures but were never seen in field specimens. Galls appeared as unorganized tissue found primarily on males and bisexuals, but occasionally on females and tetrasporophytes. The gall cells usually were less pigmented than neighboring tissue, but contained cells with fluorescent plastids and nuclei. The galls were not transferable to other potential hosts. Galls could be produced from gall-free tissue of cultures that originally had galls even after transfer to new culture dishes. Electon microscopy of galls on one isolate (3895) showed that virus-like particles are observed in some gall cells. It is possible that a virus is the causative agent of these galls.

• Journal of Plant Biotechnology
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• v.8 no.1
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• pp.15-19
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• 2006

An efficient protocol for in vitro multiple shoot bud induction and plant regeneration from mature green cotyledon derived callus tissues of Acacia sinuata has been developed. Callus formation occurs at all the concentrations of thidiazuron (TDZ) in Murashige and Skoog's (MS) medium, but 0.6 ${\mu}M$ proved to be the best with maximum callus formation frequency. Supplementation of TDZ in combination with indole-acetic acid (IAA) in MS media accelerates shoot bud organogenesis in differentiating callus tissues with 60-70% conversion of shoot buds into shoot Most efficient shoot organogenesis was recorded when TDZ induced calli were subcultured at different concentrations of 6-benzyla-denine (BA). Optimum shoot bud induction and plant regeneration from callus was achieved when 0.6 ${\mu}M$ (TDZ) induced calli were subcultured at 3.0 ${\mu}M$ (BA) where $16.6{\pm}0.74$ shoots/unit callus on obtained. Rooting in in vitro differentiated shoots was achieved when transferred to medium containing different concentration of indole-3-butyric acid (IBA) in full & half strength MS medium. The well rooted plantlets were hardened and transferred to net house with 90% survival rate.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.17 no.4
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• pp.331-336
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• 1995

To use cultured mucosa as a graft of full thickness, our laboratory has been involved in the development of techniques to grow epidermis together with connective tissue. Human oral mucosa was obtained at dental surgery. Under sterile conditions the tissues were cut into explants of 0.1 $cm^2$ which were placed in the center of 24 well tissue culture dishes and incubated in a growth medium. The growth medium used for epithelial was MEM(Minimum Essential Medium) supplemented with 10% fetal calf serum, 0.5% dimethyl sulfoxide, glutamine (0.292 g/l), epidermal growth factor (40 ug/ml), cholera toxin (30 ng/ml), hydrocortisone (2 ug/ml), insulin (40 ug/ml) and transferin (5 ug/ml). The medium for stratification of epithelial cells was MEM supplemented with 10% fetal calf serum, 0.5% dimethyl sulfoxide and glutamine (0.292 g/l). The medium used for fibroblasts was MEM supplemented with 10% fetal calf serum. With the three types of media used alternatively, a mucosa composed of epidermis and connective tissue was obtained after 3 weeks of culture.