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標題: 細胞計數法的研發與TiO2-Mica基板對細胞成長影響的探討
Development of Cell Counting and TiO2-Mica Substrate Effects on Cell Growth
作者: 陳孟宏
Chen, Meng-Hung
關鍵字: cell counting
出版社: 奈米科學研究所
引用: 1、Qian Cheng, Kyriakos Komvopoulos, Song Li. 2011. Surface chemical patterning for long-term single-cell culture. Journal of Biomedical Materials Research Part A 96A:3, 507-512. 2、Natália M. Alves, Iva Pashkuleva, Rui L. Reis, João F. Mano. 2010. Controlling Cell Behavior Through the Design of Polymer Surfaces. Small 6:20, 2208-2220. 3、Abhinav P. Acharya, Michael J. Clare-Salzler, Benjamin G. Keselowsky. 2009. A high-throughput microparticle microarray platform for dendritic cell-targeting vaccines. Biomaterials 30:25, 4168-4177. 4、Catherine A. Goubko, Xudong Cao. 2009. Patterning multiple cell types in co-cultures: A review. Materials Science and Engineering: C 29:6, 1855-1868. 5、Christopher J. Bettinger, Robert Langer, Jeffrey T. Borenstein. 2009. Engineering Substrate Topography at the Micro- andNanoscale to Control Cell Function. Angewandte Chemie International Edition 48:30, 5406-5415. 6、Adam W. Feinberg, James F. Schumacher, Anthony B. Brennan. 2009. Engineering high-density endothelial cell monolayers on soft substrates. Acta Biomaterialia 5:6, 2013-2024. 7、Joseph M. Corey, Caitlyn C. Gertz, Thomas J. Sutton, Qiaoran Chen, Katherine B. Mycek, Bor-Shuen Wang, Abbey A. Martin, Sara L. Johnson, Eva L. Feldman. 2009. Patterning N-type and S-type neuroblastoma cells with Pluronic F108 and ECM proteins. Journal of Biomedical Materials Research Part A 9999A, NA-NA. 8、Jianhua Wei, Masao Yoshinari, Shinji Takemoto, Masayuki Hattori, Eiji Kawada, Baolin Liu, Yutaka Oda. 2007. Adhesion of mouse fibroblasts on hexamethyldisiloxane surfaces with wide range of wettability. Journal of Biomedical Materials Research Part B: Applied Biomaterials 81B:1, 66-75. 9、Mauris N. De Silva, Jason Paulsen, Michael J. Renn, David J. Odde. 2006. Two-step cell patterning on planar and complex curved surfaces by precision spraying of polymers. Biotechnology and Bioengineering 93:5, 919-927. 10、Nathan J. Sniadecki, Ravi A. Desai, Sami Alom Ruiz, Christopher S. Chen. 2006. Nanotechnology for Cell–Substrate Interactions. Annals of Biomedical Engineering 34:1, 59-74. 11、2006. Selective Attachment of Mammalian Cells and Polystyrene Microbeads to Functional Amine Patterns Defined by Perfluoroalkyl Silane Surfaces. Bulletin of the Korean Chemical Society 27:1, 37-38. 12、Nathan J. Sniadecki, Ravi A. Desai, Sami Alon Ruiz, and Christopher S. Chen. Nanotechnology for Cell–Substrate Interactions. Annals of Biomedical Engineering, Vol.34, No.1, January 2006 59–74 13、Molly M. Stevens, Michael Mayer, Daniel G. Anderson, Douglas B. Weibel, George M. Whitesides, Robert Langer.2005. Direct patterning of mammalian cells onto porous tissue engineering substrates using agarose stamps. Biomaterials 26:36, 7636-7641. 14、120Tan, J. L.,W. Liu, C. M. Nelson, S. Raghavan, and C. S. Chen. Simple approach to micropattern cells on common culture substrates by tuning substrate wettability. Tissue Eng. 10:865–872, 2004. 15、Electrochemical atomic force microscopic studies on passive films of nitrogen-bearing austenitic stainless steels;U. Kamachi Mudali ,Y. Katada Electrochimica Acta 46 (2001) 3735–3742 16、In-situ electrochemical atomic force microscopy studies of aqueous corrosion and inhibition of copper;G. Bertrand,E. Rocca,C. Savall,C. Rapin,J.-C. Labrune,P. Steinmetz;Journal of Electroanalytical Chemistry 489 (2000) 38–45 17、K.-F. Giebel, C. Bechinger, S. Herminghaus, M. Riedel,P. Leiderer, U. Weiland, and M. Bastmeyer. Imaging of Cell/Substrate Contacts of Living Cells with Surface Plasmon Resonance Microscopy. Biophysical Journal Volume 76 January 1999 509–516 18、AFM tip induced selective electrochemical etching of and metal deposition on p-GaAs(100) surface;Michio Koinuma, Kohei Uosaki Surface Science 357-358 (1996) 565-570 19、Ruxiong Cai, Yoshinobu Kubota, Taro Shuin, Hideki Sakai, Kazuhito Hashimoto, and Akira Fujishima , Induction of Cytotoxicity by Photoexcited TiO2 Paiticles. Cancer Research 52.2346-2348.April 15.1992. 20、Cai, R., Hashimoto, K.. Itoh, K.. Kubota, Y., and Fujishima, A. Photokilling of malignant cells with TiO2 powder. Bull. Chem. Soc. Jpn., 64: 1268-1273, 1991. 21、Kodama. S., and Yagi, S. Photocatalytic reactions of 1,3-butadiene over water-adsorbed TiO2. J. Phys. Chem., 94: 5015-5019, 1990. 22、奈米檢測技術,國家實驗研究院儀器科技研究中心2009年出版 23、圖解奈米科技與光觸媒,商周出版社,呂宗昕著
摘要: 中文摘要 在本研究中,進行了NIH 3T3細胞在不同基板上成長情況的探討,提出了一項有效且可信的貼附性細胞計數法,並探討NIH 3T3在不同濃度TiO2的塗佈造成不同表面粗造度的mica基板上生長情形的異同。 本實驗所選用的細胞為NIH 3T3(小鼠纖維母細胞)細胞。為了比較面臨各種情況時細胞的成長情況,需要一個方便可信且不干擾細胞的貼附型細胞計數法,『NSL細胞計數法』因此被成功地發展出來了,其測試結果相當適合用來計數貼附型細胞,其正確性更已被驗證與傳統的血球板細胞計數法相當。 細胞增長的兩大機制為細胞貼附與細胞增殖,使用AFM(原子力顯微鏡)可以輕易且正確地偵測並計算基板表面的粗造度,以此方式可以清楚地發現細胞貼附的難易度與基板表面的粗造度是有關係的。而基板的表面活性則明顯地影響了接下來7天的細胞增值情形。在不同濃度TiO2的塗佈造成不同表面粗造度的mica基板上進行研究,發現了粗造的表面較有利於細胞的貼附,同時進行了針對奈米TiO2顆粒照射紫外光的研究,發現因此而產生的自由基O2-與•OH嚴重地影響了細胞的增殖。 總結來說,論文主體分為三大部分,安排如下。在第2章中,介紹了mica基板的製作以及TiO2的塗佈法。第3章則介紹了本研究中所使用的細胞培養技術。第4章則是『NSL細胞計數法』的研發過程以及各項研究結果與討論。最後則是未來展望。
Abstract In this work, the proliferation of NIH-3T3 cells on various substrates was investigated. A development of surface-supported cell counting was brought up with a novel idea. The influence of surface roughness on cell attachment with nano-particle was also studied. Herein, NIH-3T3 cells (mouse embryo fibroblast cell) were chosen as testing system. A surface-supported counting, which does not disturb the cell proliferation, was first proposed. The reliability and critical comparisons of various methods for cell counting were studied. The result showed that our NSL cell counting was suitable for Anchorrage-dependent cell investigation. The mechanisms of cell proliferation are controlled by two steps: cell attachment and cell proliferation. The surface roughness, which was examined using Atomic Force Microscope (AFM), was found to be responsible for cell attachment. The surface activity was referred to the population of cells after 7-day investigation. The TiO2 nano-particles here served as the surface roughcast. The result shows that cell attachments enhanced with surface roughness. Moreover, the UV irradiation for TiO2 nanoparticles was applied before and after cell culture. The presence of O2- and •OH was in the charge of cell proliferation. In summary, the thesis is divided into 3 sections arranged in a manner that follows the procedures of cell counting on various substrates. In the chapter 2, introductions to mica and TiO2 nano-particles were brought up. In the chapter 3, the cell line culture on various substrates was adapted. The chapter 4 was included the experimental results and discussion. Finally, the future work was proposed to improve the fabrication of bio-scaffolds.
其他識別: U0005-1401201209382500
Appears in Collections:奈米科學研究所



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