請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/81796完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 許聿翔(Yu-Hsiang Hsu) | |
| dc.contributor.author | Hong-Wen Wang | en |
| dc.contributor.author | 王泓文 | zh_TW |
| dc.date.accessioned | 2022-11-25T03:03:47Z | - |
| dc.date.available | 2023-08-11 | |
| dc.date.copyright | 2021-11-09 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-08-11 | |
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Xiao et al., 'Microfabricated perfusable cardiac biowire: a platform that mimics native cardiac bundle,' Lab on a Chip, vol. 14, no. 5, pp. 869-882, 2014. [13] N. Tandon, A. Marsano, R. Maidhof, L. Wan, H. Park, and G. Vunjak‐Novakovic, 'Optimization of electrical stimulation parameters for cardiac tissue engineering,' Journal of tissue engineering and regenerative medicine, vol. 5, no. 6, pp. e115-e125, 2011. [14] C.-H. Chan, 'Development of Aligned P(VDF-TrFE) Piezoelectric Nanofiber Bundles for Cardiac Drug Screening Application.,' Master's Thesis of Institute of Applied Mechanics. National Taiwan University, Taiwan (R.O.C) 2017). [15] Y.-C. Chan et al., 'Electrical stimulation promotes maturation of cardiomyocytes derived from human embryonic stem cells,' Journal of cardiovascular translational research, vol. 6, no. 6, pp. 989-999, 2013. [16] E. Serena et al., 'Electrical stimulation of human embryonic stem cells: cardiac differentiation and the generation of reactive oxygen species,' Experimental cell research, vol. 315, no. 20, pp. 3611-3619, 2009. [17] A. Brevet, E. Pinto, J. Peacock, and F. E. Stockdale, 'Myosin synthesis increased by electrical stimulation of skeletal muscle cell cultures,' Science, vol. 193, no. 4258, pp. 1152-1154, 1976. [18] P. M. McDonough and C. Glembotski, 'Induction of atrial natriuretic factor and myosin light chain-2 gene expression in cultured ventricular myocytes by electrical stimulation of contraction,' Journal of Biological Chemistry, vol. 267, no. 17, pp. 11665-11668, 1992. [19] Y. Xia, L. M. Buja, R. C. Scarpulla, and J. B. McMillin, 'Electrical stimulation of neonatal cardiomyocytes results in the sequential activation of nuclear genes governing mitochondrial proliferation and differentiation,' Proceedings of the National Academy of Sciences, vol. 94, no. 21, pp. 11399-11404, 1997. [20] W.-H. Zimmermann et al., 'Tissue engineering of a differentiated cardiac muscle construct,' Circulation research, vol. 90, no. 2, pp. 223-230, 2002. [21] W. Dou et al., 'A microdevice platform for characterizing the effect of mechanical strain magnitudes on the maturation of iPSC-Cardiomyocytes,' Biosensors and Bioelectronics, vol. 175, p. 112875, 2021. [22] D. Carson et al., 'Nanotopography-induced structural anisotropy and sarcomere development in human cardiomyocytes derived from induced pluripotent stem cells,' ACS applied materials interfaces, vol. 8, no. 34, pp. 21923-21932, 2016. [23] J. R. Macadangdang et al., 'Engineered developmental niche enables predictive phenotypic screening in human dystrophic cardiomyopathy,' bioRxiv, p. 456301, 2018. [24] J. Kim et al., 'Quantitative evaluation of cardiomyocyte contractility in a 3D microenvironment,' Journal of biomechanics, vol. 41, no. 11, pp. 2396-2401, 2008. [25] J. U. Lind et al., 'Instrumented cardiac microphysiological devices via multimaterial three-dimensional printing,' Nature materials, vol. 16, no. 3, pp. 303-308, 2017. [26] N. Bursac, K. Parker, S. Iravanian, and L. Tung, 'Cardiomyocyte cultures with controlled macroscopic anisotropy: a model for functional electrophysiological studies of cardiac muscle,' Circulation research, vol. 91, no. 12, pp. e45-e54, 2002. [27] A. Agarwal, J. A. Goss, A. Cho, M. L. McCain, and K. K. Parker, 'Microfluidic heart on a chip for higher throughput pharmacological studies,' Lab on a Chip, vol. 13, no. 18, pp. 3599-3608, 2013. [28] I. Mannhardt et al., 'Human engineered heart tissue: analysis of contractile force,' Stem cell reports, vol. 7, no. 1, pp. 29-42, 2016. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/81796 | - |
| dc.description.abstract | 現今人類誘導多能幹細胞（HiPSCs）的開發和所開發的心肌細胞(hiPSC-CMs)分化技術是再生醫學和心血管的主要研究關鍵，但由於這些細胞是誘導分化而成，皆尚未成熟，若沒有像體內的生理環境刺激，將會影響後續篩藥及實驗結果的準確性，因此，如何提高心肌細胞成熟度成為重要的研究重點。在心臟藥物開發階段，需要耗費大量的時間及成本進行藥物篩檢，而現今商業所使用的多孔盤多為塑膠材料製成，雖然使用多孔盤進行大量且自動化篩檢的技術已非常成熟，但其底部為塑膠材質，材料硬度約2GPa左右，且厚度在1mm到0.5mm 之間，遠高於體內生理環境硬度，使在塑膠多孔盤上培養的心肌細胞發展較不成熟，與體內的生理訊表現有差異，將造成藥物篩選結果的失準。而目前常用於學界的材料為PDMS矽膠，其為硬度約3MPa的高柔性材料，較適合用來培養細胞及進行實驗，但PDMS缺點為製造價格昂貴，且不易大量生產製造及發展成商品化的產品。因此，本研究開發出以塑膠薄膜為基材，利用薄膜低撓曲剛度的機械性質，使其可以在心肌細胞收縮時產生彎曲形變，用以補償塑膠材料高於心肌細胞體內環境硬度約5個數量級的差異，可提供量產且更近似體內環境的全塑膠基材來培養心肌細胞。 為探討以降低膜厚來有效提升薄膜的柔性，本研究比較心肌細胞培養在75μm至5μm膜厚上的成熟度表現，並以理論推導及有限元素模擬驗證其柔性提升的效果，證明薄膜厚度對撓曲剛度及形變量的影響可達3個數量級，本研究並在薄膜上加入微溝槽結構，以再次降低薄膜的撓曲剛度並誘導心肌細胞排列，以促使其成熟。由實驗結果得知，當平面薄膜基材厚度由75μm下降至5μm，肌節長度平均增加了9.14%，而在單一心肌細胞培養條件下，肌節長度增加了7.05%，驗證了撓曲剛度及形變量對心肌細胞成熟度之影響。在基材加入微結構後，可以發現肌節長度在任何培養參數下，皆有明顯的增加，在75μm薄膜上提升6.04%，且培養於微溝槽上的心肌細胞在有電刺激的狀況下，其成熟度與沒有電刺激的組別沒有顯著的差異，證明以具有微溝槽的薄膜培養心肌細胞，可提供適當的基材機械性質來培養心肌細胞。本研究成功將薄膜塑膠基材轉化變為適合心肌細胞生長的高柔性基材，並結合的微溝槽結構達到提升細胞排列及成熟度的效果，未來將可能結合塑膠孔盤進行商業化製造，達到符合大量藥物篩檢的目標。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-25T03:03:47Z (GMT). No. of bitstreams: 1 U0001-1108202114355300.pdf: 5044138 bytes, checksum: f7e3995ce61f8e0e42944ca2ab965a75 (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | "口試委員會審定書 i 誌謝 ii 摘要 iii ABSTACT iv 目錄 vi 圖目錄 ix 表目錄 xiii 第1章 緒論 1 1.1 誘導多能幹細胞 (iPSC)介紹 1 1.2 研究背景及動機 1 1.3 研究目的 3 1.4 論文架構 4 第2章 文獻回顧 5 2.1 心肌細胞之性質及特性 5 2.2 促進心肌細胞成熟之方法 5 2.2.1 電生理刺激 5 2.2.2 機械刺激 7 2.2.3 基材幾何結構 8 2.2.4 細胞外基質 9 2.2.5 基材硬度 9 第3章 設計理念 10 3.1 心肌細胞培養平台設計 12 3.1.1 細胞外基質(Extracellular Matrix, ECM) 12 3.1.2 細胞培養槽(Culture well) 12 3.1.3 細胞濃度設計 12 3.2 厚度及微結構凹槽之薄膜基材設計 13 3.3 外部電生理刺激系統 14 第4章 實驗方法 15 4.1 薄膜基材理論推導 15 4.1.1 中心軸及截面二次矩公式推導 15 4.1.2 曲率半徑公式推導 18 4.1.3 薄膜位移量計算 21 4.2 微結構晶片製程 24 4.2.1 光罩繪製及製造 24 4.2.2 黃光微影製程 25 4.2.3 晶片微結構量測 28 4.3 微結構塑膠薄模之熱壓印製程 29 4.3.1 熱壓印製程 29 4.3.2 聚丙烯模之微結構量測 31 4.4 電刺激生理系統 33 4.4.1 電刺激參數 33 4.4.2 電極設計 34 4.5 心肌細胞之培養 35 4.5.1 心肌細胞培養液 35 4.5.2 細胞外基質 36 4.5.3 心肌細胞解凍 36 4.5.4 心肌細胞培養 36 4.5.5 心肌細胞培養流程 37 4.6 心肌細胞染色、成像、定量及ANOVA分析 37 4.6.1 細胞固定 37 4.6.2 螢光染色步驟 38 4.6.3 光學成像系統 38 4.6.4 心肌細胞定量 39 4.6.5 ANOVA數據分析 40 第5章 有限元素模擬分析 42 5.1 有限元素模型之建立與參數設定 42 5.2 心肌細胞收縮對不同厚度之具微結構基材影響之模擬 44 第6章 實驗結果與討論 46 6.1 薄膜之理論推導及有限元素法模擬分析結果 46 6.1.1 薄膜理論推導結果 46 6.1.2 具微結構薄膜之有限元素法模擬分析結果 47 6.1.3 理論推導及有限元素法模擬分析結果比較 49 6.2 不同厚度之平面塑膠薄膜基材對心肌細胞之影響 50 6.2.1 不同厚度平面塑膠薄膜基材對高濃度培養心肌細胞之影響 50 6.2.2 不同厚度平面塑膠薄膜基材對低濃度培養心肌細胞之影響 51 6.3 微結構對心肌細胞之影響 55 6.3.1 微結構塑膠薄膜基材對高濃度培養心肌細胞之影響 55 6.3.2 微結構塑膠薄膜基材對低濃度培養心肌細胞之影響 57 6.4 外部電刺激對培養於平面基材之心肌細胞之影響 60 6.4.1 電刺激對培養於微結構基材之高濃度心肌細胞之影響 60 6.4.2 電刺激對培養於平面基材之低濃度心肌細胞之影響 61 6.4.3 電刺激對培養於微結構基材之低濃度心肌細胞之影響 65 6.5 細胞濃度差異對心肌細胞之影響 68 6.6 總比較 69 6.6.1 高細胞濃度總比較 69 6.6.2 低細胞濃度總比較 70 第7章 討論、結論及未來展望 73 7.1 討論 73 7.2 結論 74 7.3 未來展望 75 REFERENCES 76 " | |
| dc.language.iso | zh-TW | |
| dc.subject | 實驗室晶片 | zh_TW |
| dc.subject | 心肌細胞 | zh_TW |
| dc.subject | 撓曲剛度 | zh_TW |
| dc.subject | 薄膜晶片 | zh_TW |
| dc.subject | 微溝槽結構 | zh_TW |
| dc.subject | lab on a chip | en |
| dc.subject | cardiomyocyte | en |
| dc.subject | microgroove | en |
| dc.subject | flexural rigidity | en |
| dc.subject | thin- film device | en |
| dc.title | 撓曲剛度及微溝槽結構對心肌細胞成熟度影響之研究 | zh_TW |
| dc.title | Study on the influence of the flexural rigidity and microgrooves on the maturation of cardiomyocytes | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 謝清河(Hsin-Tsai Liu),游佳欣(Chih-Yang Tseng),黃瀞瑩 | |
| dc.subject.keyword | 心肌細胞,微溝槽結構,撓曲剛度,薄膜晶片,實驗室晶片, | zh_TW |
| dc.subject.keyword | cardiomyocyte,microgroove,flexural rigidity,thin- film device,lab on a chip, | en |
| dc.relation.page | 77 | |
| dc.identifier.doi | 10.6342/NTU202102276 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2021-08-13 | |
| dc.contributor.author-college | 工學院 | zh_TW |
| dc.contributor.author-dept | 應用力學研究所 | zh_TW |
| dc.date.embargo-lift | 2023-08-11 | - |
| 顯示於系所單位： | 應用力學研究所 | |
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| U0001-1108202114355300.pdf | 4.93 MB | Adobe PDF | 檢視/開啟 |
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