• Title/Summary/Keyword: Three-dimensional culture

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Investigating the Morphology and Kinetics of Three-Dimensional Neuronal Networks on Electro-Spun Microstructured Scaffolds

  • Kim, Dongyoon;Kim, Seong-Min;Kang, Donghee;Baek, Goeun;Yoon, Myung-Han
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.277.2-277.2
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    • 2013
  • Petri dishes and glass slides have been widely used as general substrates for in vitro mammalian cell cultures due to their culture viability, optical transparency, experimental convenience, and relatively low cost. Despite the aforementioned benefit, however, the flat two-dimensional substrates exhibit limited capability in terms of realistically mimicking cellular polarization, intercellular interaction, and differentiation in the non-physiological culture environment. Here, we report a protocol of culturing embryonic rat hippocampal neurons on the electro-spun polymeric network and the results from examination of neuronal cell behavior and network formation on this culture platform. A combinatorial method of laser-scanning confocal fluorescence microscopy and live-cell imaging technique was employed to track axonal outgrowth and synaptic connectivity of the neuronal cells deposited on this model culture environment. The present microfiber-based scaffold supports the prolonged viability of three-dimensionally-formed neuronal networks and their microscopic geometric parameters (i.e., microfiber diameter) strongly influence the axonal outgrowth and synaptic connection pattern. These results implies that electro-spun fiber scaffolds with fine control over surface chemistry and nano/microscopic geometry may be used as an economic and general platform for three-dimensional mammalian culture systems, particularly, neuronal lineage and other network forming cell lines.

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The Differentiation of bone Marrow Stromal Cells into NP-like Cells through 3-Dimensional Co-culture System (3차원 Co-culture 시스템을 통한 BMSC의 NP-like Cell로의 분화)

  • Kim, D.H.;Kim, S.H.;Heo, S.J.;Shin, J.W.;Kim, Y.J.;Park, S.H.;Jun, J.W.;Shin, J.W.
    • Journal of Biomedical Engineering Research
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    • v.29 no.2
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    • pp.159-163
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    • 2008
  • The goal of this study is to investigate the effect and potential of three-dimensional Co-culture of BMSCs (bone marrow stromal Cells) and NP (nucleus pulposus) Cells on the differentiation of BMSCs into NP-like Cells. The NP Cells and BMSCs were isolated and cultured from New Zealand White rabbits. The isolated NP Cells and BMSCs were prepared in different alginate beads. Those two types of beads were separated by a track-etched membrane of $3\;{\mu}m$ pore in a 6-well culture plate. No growth factors were used. In addition to these, NP and BMSC were cultured in the beads independently for control. The number of Cells in Co-culturing system was half of those in two control groups. Proliferation and production of glycosaminoglycan (GAG) were evaluated along with histological observation. The GAG production rate(GAG contents/Cell) of Co-cultured BMSCs were much higher than that of BMSCs cultured alone. The total amounts of GAG produced by BMSCs in Co-culturing system were larger than those produced by BMSCs in control group and were comparable with those produced by NP alone even the number of each Cell was half of BMSCs in Co-culturing system. This study showed the potential of differentiation of BMSCs into NP-like Cells through three-dimensional Co-culture system even without any chemical agents.

Identification of matrix metalloproteinases secreted by human hepatocarcinoma HepG2 cells

  • Lee, Young Jae;Kim, Keun Cheon;Lim, Jeong Mook;Lee, Seung Tae
    • Journal of Animal Reproduction and Biotechnology
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    • v.37 no.1
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    • pp.62-66
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    • 2022
  • To date, the development of anticancer drugs has been conducted using two-dimensional (2D) cell culture systems. However, since cancer cells in the body are generated and developed in three-dimensional (3D) microenvironments, the use of 2D anticancer drug screening can make it difficult to accurately evaluate the anticancer effects of drug candidates. Therefore, as a step towards developing a cancer cell-friendly 3D microenvironment based on a combination of vinylsulfone-functionalized polyethylene glycol (PEG-VS) with dicysteine-containing crosslinker peptides with an intervening matrix metalloproteinase (MMP)-specific cleavage site, the types of MMPs secreted from human hepatocarcinoma HepG2 cells, a representative cancer cell, were analyzed transcriptionally and translationally. MMP3 was confirmed to be the most highly expressed protease secreted by HepG2 cells. This knowledge will be important in the design of a crosslinker necessary for the construction of PEG-based hydrogels customized for the 3D culture of HepG2 cells.

'Barrage' phenomenon in Chinese Two-dimensional cultural environment (중국 2차원문화 환경에서의 '탄막(弹幕)'현상)

  • Shi, Xianglan;Choi, eunkyoung
    • Cartoon and Animation Studies
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    • s.50
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    • pp.23-42
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    • 2018
  • Currently, two-dimensional culture is the main keyword in Chinese contents market. In China, many capital and economic giants have entered into two-dimensional cultural market because the number of users and the interest of the public for two-dimensional culture are increasing day by day. The reason for the rise of the two-dimensional culture into the mainstream culture is not only the switching of the mainstream consumer class but also Two-Dimensional Culture forms a huge industrial chain covering many areas including movies, performances and theme parks in addition to ACGN (Animation, Comic, Game and light Novel), the second is that more people are participating in Two-Dimensional Culture due to the specific audience with high social environment and loyalty and the third is looking for the role of the users in Two-Dimensional cultural content from three characteristics, such as strong cultural prevalence. This study analyzed the Chinese two-dimensional cultural environment through PEST(Political, Economic, Social and Technological analysis) In addition, as the ecological environment and communication method of Two-Dimensional Culture users changed, the users became active participants and the second producers of contents. I examined the way of using the users' contents through the phenomenon of 'Barrage'. Therefore, this study discussed the appropriate development path for the user-oriented contents industry in the Chinese Two-Dimensional cultural environment.

A review on three dimensional scaffolds for tumor engineering

  • Ceylan, Seda;Bolgen, Nimet
    • 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.

Efficacy of biological inhibitors in three-dimensional culture models of oral squamous cell carcinoma

  • Eun Kyoung Kim;Sook Moon;Myung-Jin Lee;Dokyeong Kim
    • International Journal of Oral Biology
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    • v.49 no.1
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    • pp.18-25
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    • 2024
  • Despite advancements in therapeutic approaches, radiotherapy and cisplatin-based chemotherapy remain primary noninvasive treatments for patients with oral squamous cell carcinoma (OSCC). Moreover, the 5-year survival rate for patients with OSCC has remained almost unchanged for several decades, and many side effects of chemotherapy still exist. In this study, three-dimensional (3D) models of OSCC were established using fibroblasts, and the efficacy of various biological inhibitors was evaluated. A culture of epithelial cells with two types of fibroblasts (hTERT-hNOFs and cancer-associated fibroblasts) within a type I collagen matrix resulted in the formation of a continuous layer of tightly packed cells compared to models without fibroblasts. Furthermore, the effects of biological chemicals, including Y27632, latrunculin A, and verteporfin, on these models were investigated. The stratified formation of the epithelial layer and invasion in OSCC 3D-culture models were effectively inhibited by verteporfin, whereas invasion was weakly inhibited by Y27632 and latrunculin. Collectively, the developed OSCC 3D-culture models established with fibroblasts demonstrated the potential for drug screening, with verteporfin showing promising efficacy.

Expression types and aesthetic characteristics of modern fashion applying the formativeness of symmography (시모그래피의 조형성을 응용한 현대 패션 디자인의 표현유형과 미적 특성 연구)

  • Kwon, Giyoung
    • The Research Journal of the Costume Culture
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    • v.29 no.3
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    • pp.361-373
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    • 2021
  • The purpose of this study is to contribute to the role of lines in creative design development by analyzing the expression types and aesthetic characteristics of modern fashion using geometric formativeness of symmography. A literature study was conducted of works since 2009 to examine the general consideration of lines together with analysis of the concept and characteristics of symmography in the formative arts field, and to analyze the expression types and aesthetic characteristics of modern fashion design using the formativeness of symmography. The infinite sense of formativeness and original expression of symmography are used in formative arts such as space design, installation art, and industrial design. Expression types of modern fashion design using geometric formativeness of symmography can be classified into the following three types: two-dimensional graphic pattern, relief surface, and three-dimensional spatial. First, the two-dimensional graphic pattern type forms an optical pattern, providing individuality and visual interest to the textile design. Second, the relief surface type expresses the plane in various ways, so that the thickness changes according to how lines overlap. Third, the three-dimensional spatial type expands the boundaries of clothing and creates a fantastic spatial beauty. Next, the aesthetic formativeness of fashion design using symmography can be classified into repetitive rhythmicity, geometric self-similarity, and optical spatiality. Symmography enables a myriad of geometric patterns to be developed depending on material, color, and the designer's imagination, and helps inspire a variety of designs in fashion that sculpt a three-dimensional human body.

PERIPHERAL NERVE REGENERATION USING A THREE-DIMENSIONALLY CULTURED SCHWANN CELL CONDUIT (삼차원 배양된 슈반세포 도관을 이용한 말초 신경 재생)

  • Kim, Soung-Min;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.30 no.1
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    • pp.1-16
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    • 2004
  • The use of artificial nerve conduit containing viable Schwann cells is one of the most promising strategies to repair the peripheral nerve injury. To fabricate an effective nerve conduit whose microstructure and internal environment are more favorable in the nerve regeneration than existing ones, a new three-dimensional Schwann cell culture technique using $Matrigel^{(R)}$. and dorsal root ganglion (DRG) was developed. Nerve conduit of three-dimensionally arranged Schwann cells was fabricated using direct seeding of freshly harvested DRG into a $Matrigel^{(R)}$ filled silicone tube (I.D. 1.98 mm, 14 mm length) and in vitro rafting culture for 2 weeks. The nerve regeneration efficacy of three-dimensionally cultured Schwann cell conduit (3D conduit group, n=6) was assessed using SD rat sciatic nerve defect of 10 mm, and compared with that of silicone conduit filled with $Matrigel^{(R)}$ and Schwann cells prepared from the conventional plain culture method (2D conduit group, n=6). After 12 weeks, sciatic function was evaluated with sciatic function index (SFI) and gait analysis, and histomorphology of nerve conduit and the innervated tissues of sciatic nerve were examined using image analyzer and electromicroscopic methods. The SFI and ankle stance angle (ASA) in the functional evaluation were $-60.1{\pm}13.9$, $37.9^{\circ}{\pm}5.4^{\circ}$ in 3D conduit group (n=5) and $-87.0{\pm}12.9$, $32.2^{\circ}{\pm}4.8^{\circ}$ in 2D conduit group (n=4), respectively. And the myelinated axon was $44.91%{\pm}0.13%$ in 3D conduit group and $13.05%{\pm}1.95%$ in 2D conduit group to the sham group. In the TEM study, 3D conduit group showed more abundant myelinated nerve fibers with well organized and thickened extracellular collagen than 2D conduit group, and gastrocnemius muscle and biceps femoris tendon in 3D conduit group were less atrophied and showed decreased fibrosis with less fatty infiltration than 2D conduit group. In conclusion, new three-dimensional Schwann cell culture technique was established, and nerve conduit fabricated using this technique showed much improved nerve regeneration capacity than the silicone tube filled with $Matrigel^{(R)}$ and Schwann cells prepared from the conventional plain culture method.

Evaluation of polyglycolic acid as an animal-free biomaterial for three-dimensional culture of human endometrial cells

  • Sadegh Amiri;Zohreh Bagher;Azadeh Akbari Sene;Reza Aflatoonian;Mehdi Mehdizadeh;Peiman Broki Milan;Leila Ghazizadeh;Mahnaz Ashrafi;FatemehSadat Amjadi
    • Clinical and Experimental Reproductive Medicine
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    • v.49 no.4
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    • pp.259-269
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    • 2022
  • Objective: Animal-free scaffolds have emerged as a potential foundation for consistent, chemically defined, and low-cost materials. Because of its good potential for high biocompatibility with reproductive tissues and well-characterized scaffold design, we investigated whether polyglycolic acid (PGA) could be used as an animal-free scaffold instead of natural fibrin-agarose, which has been used successfully for three-dimensional human endometrial cell culture. Methods: Isolated primary endometrial cells was cultured on fibrin-agarose and PGA polymers and evaluated various design parameters, such as scaffold porosity and mean fiber diameter. Cytotoxicity, scanning electron microscopy (SEM), and immunostaining experiments were conducted to examine cell activity on fabricated scaffolds. Results: The MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay and SEM results showed that endometrial cells grew and proliferated on both scaffolds. Immunostaining showed cytokeratin and vimentin expression in seeded cells after 7 days of culture. On both scaffolds, an epithelial arrangement of cultured cells was found on the top layer and stromal arrangement matrix on the bottom layer of the scaffolds. Therefore, fibrin-agarose and PGA scaffolds successfully mimicked the human endometrium in a way suitable for in vitro analysis. Conclusion: Both fibrin-agarose and PGA scaffolds could be used to simulate endometrial structures. However, because of environmental and ethical concerns and the low cost of synthetic polymers, we recommend using PGA as a synthetic polymer for scaffolding in research instead of natural biomaterials.

Colors of Costume in Korean Basic Culture

  • Kim, Ji-Young;Kim, Young-In
    • Proceedings of the Korea Society of Costume Conference
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    • 2003.10a
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    • pp.32-32
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    • 2003
  • This research has for its object to look carefully into the peculiarity of the color of the costume in the Korean culture by revealing the characteristic and the idea of the color being discovered in the Korean basic culture, which has the majority of the Korean people. The scope of the basic culture was divided into folk belief, folk game and folk play. Within these limits, the colors of the dress, accessories, instruments were extracted by comparing with the naked eye in NCS Color System. For the analysis of hue and tone the secondary dimensional analysis using NCS color system and the three-dimensional analysis using the software, COLOR 3D Version 2.0, were done.

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