博碩士論文 111324041 詳細資訊



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姓名 蔡弘源(HUNG-YUAN TSAI)  查詢紙本館藏   畢業系所 化學工程與材料工程學系
論文名稱 利用組織工程製作仿生腫瘤 並以MRI進行影像分析
(Preparation of A Bionic Tumor using Tissue Engineering and Its Image Analysis by MRI)
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摘要(中) 本研究針對不同腫瘤細胞接種方法，在三維明膠/羥基磷灰石/戊二醛（GHG）支架中所形成腫瘤仿體。利用被動接種的蓋壓法，以及主動接種的培養液注射法和明膠注射法將細胞植入支架中，並通過磁共振成像（MRI）觀察腫瘤組織的生長情況。結果顯示，蓋壓法會導致細胞主要集中於細胞液接觸面；培養液注射法中，細胞能分布於支架的注射上層與注射層，而注射下層的細胞液可能因重力作用而流失。明膠注射法中，觀察到支架的注射上層、注射層與注射下層的孔洞內皆有細胞被明膠包覆，並推測黏性的明膠能將細胞維持在孔洞內進行懸浮生長。在14天的培養後，三種接種方法在表層均能培養出緻密的組織團塊，其中明膠注射法不僅可以增加在組織團塊數，也能促進在支架縱向的分布。在MRI觀察中，影像解析度的限制為83 μm，因此無法成像較小的腫瘤組織或孔洞，但MRI能對腫瘤組織在支架的分布進行大尺度的觀察。此外，MRI影像能進行三維重建，但支架結構與腫瘤組織的訊號存在重疊，導致兩者無法清晰區分。未來研究可考慮對MRI影像進行優化處理，並對仿體進行連續觀察。
摘要(英) This study investigates the impact of different cell seeding methods on the formation of bionic tumors in a three-dimensional GHG（gelatin/hydroxyapatite/glutaraldehyde） scaffold. Three seeding methods were evaluated including passive seeding by capping and active seeding via culture medium injection and gelatin injection. Capping method allowed cell suspension passively absorbed to the scaffold, leading to cell primarily concentrated on the cell suspension-contacting surface. In the culture medium injection method, cells were distributed in the injection layer and its upper. Cell was unfindable in the region below the injection layer due to possible lost through gravity. In contrast, with the gelatin injection method, cells were observed encapsulated by gelatin within the pores not only in the injection layer but also its upper and lower regions, suggesting that gelatin′s supportive properties enabled cell suspended growth within the pores. After cultivation for 2 weeks, all seeding methods resulted in the formation of dense tissue, with a significant increase in vertical growth area observed in the gelatin injection method, indicating that the suspension culture in gelatin could promote tissue growth in the vertical dimension. These bionic tumor were monitored using magnetic resonance imaging（MRI）analysis.Due to the resolution limitation of 83 μm of MRI, it was difficult to image smaller tumor tissues or pores within the scaffold. However, MRI could conduct large-scale observations of the distribution of tumor tissue in the scaffold. The 3D reconstruction results of MRI showed that the signals of the scaffold structure and the tumor tissue overlaped, preventing clear distinction between the two. Future research could focus on optimizing MRI imaging and continuously monitoring tissue growth in the scaffold.
關鍵字(中) ★ 組織工程
★ 核磁共振影像		 關鍵字(英)
論文目次 摘要 I
Abstract II
誌謝 III
目錄 IV
圖目錄 V
表目錄 VI
第一章、緒論 1
1.1 現今概況 1
1.2 研究動機與挑戰 2
1.3 研究方法與目的 3
第二章、理論基礎 4
2.1癌症骨轉移 4
2.1.1 臨床上骨轉移的治療 5
2.1.2癌症骨轉移機制 6
2.1.3表皮生長因子對於骨轉移的影響 7
2.2動物模型 9
2.3 腫瘤組織體外培養 11
2.3.1 二維培養 11
2.3.2 腫瘤的細胞外基質 12
2.3.3 體外研究腫瘤大小 13
2.4 組織工程 15
2.4.1仿骨支架 16
2.4.2 體外支架材料 17
2.4.3 體外支架的細胞接種方式 19
2.4.4細胞維持在三維孔洞 21
2.4.5 體外三維仿體的培養 23
2.5腫瘤觀察方式 24
2.5.1侵入式觀察 26
2.5.2非侵入式觀察 28
2.5.2.1 MRI及MRI成像原理 30
第三章、材料與方法 32
3.1實驗藥品 32
3.2實驗儀器 33
3.3實驗流程圖 34
3.4胰臟癌細胞 35
3.4.1 胰臟癌細胞選擇與來源 35
3.4.2胰臟癌細胞繼代 36
3.5體外胰臟癌仿體 38
3.5.1仿骨支架製備 38
3.5.2製備骨轉移微環境 43
3.6胰臟癌與仿骨支架培養 48
3.6.1支架接種胰臟癌細胞 48
3.6.2計算三維仿體上的細胞數量 50
3.6.3三維動態培養系統 51
3.6.4仿體動態培養系統 53
3.7胰臟癌骨轉移仿體分析 54
3.7.1細胞假體 54
3.7.2 MRI 55
3.7.3 MRI影像分析/處理 56
3.7.4組織切片分析 61
第四章、 結果 63
4.1三維仿骨支架之特性 63
4.1.1三維仿骨支架結構與孔徑 63
4.2細胞接種 66
4.2.1蓋壓法 67
4.2.1.1蓋壓法0天的細胞垂直分布 67
4.2.1.2蓋壓法0天的細胞水平面分布 70
4.2.2 培養液注射法 71
4.2.2.1 培養液注射法0天的細胞矢狀分布 71
4.2.2.2 培養液注射法0天的細胞水平面分布 74
4.2.3 明膠注射法 75
4.2.3.1 明膠注射法0天的細胞垂直分布 75
4.2.3.2 明膠注射法0天的細胞水平面分布 78
4.3 腫瘤細胞於三維支架培養14天 80
4.3.1 蓋壓法 80
4.3.1.1蓋壓法培養後細胞的矢狀生長 80
4.3.1.2 蓋壓法培養後細胞的水平面生長 83
4.3.2 培養液注射法 85
4.3.2.1 培養液注射法培養後細胞的矢狀生長 85
4.3.2.2 培養液注射法培養後細胞的水平面生長 88
4.3.3 明膠注射法 89
4.3.3.1 明膠注射法培養後細胞的矢狀生長 89
4.3.3.2 明膠注射法培養後細胞的水平生長 93
4.3.4不同注射方法對於組織生長的比較 95
4.4 MRI的觀察 96
4.4.1 MRI平面影像上觀察組織 97
4.4.2 MRI二維影像中組織可視化提升 99
4.4.3 組織切片與MRI比較 101
4.4.4 MRI三維影像重建 105
第五章、 結論 110
第六章、 展望 112
第七章、 參考文獻 114
第八章、附錄 120
參與研討會與發表 120
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指導教授 胡威文 董國忠(Wei-Wen Hu Guo-Chung Dong) 審核日期 2024-9-20
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