微流體系統應用於機械力刺激人體膀胱癌細胞之研究

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姓名

葉立強(YE,LI-CIANG) 查詢紙本館藏

畢業系所

機械工程學系

論文名稱

微流體系統應用於機械力刺激人體膀胱癌細胞之研究

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摘要(中)

摘要
本研究的目的是設計一個機械力刺激人體膀胱癌細胞的微流體晶片，機械力包括了剪應力與正向應力。微流體晶片上有三條流道，每條流道分別有3個培養區做細胞培養，透過Syringe pump和閥門的控制分別對每條流道施加三種剪應力，再對培養區上方施加正向應力。
在對人體膀胱癌細胞刺激之前，分別對裝置中的機械力做裝置上的測試，剪應力測試使用一般染色液檢測閥門的緊密度，確保每條流道只受到單一剪應力；正向應力測試使用螢光染液觀察培養區螢光強度的變化，用來檢查培養區不會因受到外力而變形。
最後，我們將人體膀胱癌細胞(TCC)培養在微流體晶片的培養區中，分別對細胞施加3組剪應力刺激10分鐘(0.0018 dyne/cm2、0.044 dyne/cm2、0.66 dyne/cm2。)，再對每組施加3組正向應力刺激1個小時(0KPa、10KPa、100KPa)，之後在培養箱中培養24小時，對每組受到不同刺激參數細胞進行拍照，比較刺激前與刺激後的細胞數目。實驗結果顯示受到最大剪應力刺激的細胞，細胞數目有減少的趨勢；0KPa與10KPa正向應力刺激對細胞數量呈現正成長，到了100KPa細胞數目呈現了負成長。

摘要(英)

Abstract
The purpose of this study is to design a mechanical force to stimulate human bladder cancer cell microfluidic chips, mechanical forces, including the shear stress and the normal stress. The microfluidic chip has three channel, each channel has three cultures area were to do in cell culture, three shear stress is applied to each flow path through Syringe pump and control valve, respectively. And then application of normal stress on cultures area.
Human bladder carcinoma (TCC) cultured in the area of the microfluidic chip, the cells were applied to three kinds of shear stress stimulation 10 minutes (0.0018 dyne / cm2,0.044 dyne / cm2,0.66 dyne / cm2.), and then the each of three shear stress applied normal stress stimulation one hour (0KPa, 10KPa, 100KPa), then cultured in an incubator for 24 hours, subject to different stimulation parameters for each group cells were photographed, the number of stimulation cells before and after.
Experimental results show that by the cell, the cell number of the maximum shear stress stimulation downward trend; 0KPa and 10KPa normal stress stimulation on the number showed positive growth, the number of cells to 100KPa presented negative growth.

關鍵字(中)

★ 微流體晶片

關鍵字(英)

論文目次

目錄
摘要 I
Abstract II
目錄 III
圖目錄 VII
表目錄 X
第一章緒論 1
1-1前言 1
1-2文獻回顧 2
1-3研究動機及目的 9
1-4論文架構 10
第二章實驗設計與方法 11
2-1材料選擇 11
2-2微流體晶片製程 13
2-2-1黃光製程 14
2-2-2光罩製作 16
2-2-3氣閥層母模製作(負光阻) 17
2-2-4流道層母模製作(正光阻) 21
2-2-5 PDMS翻模 24
2-2-6晶片封裝 26
2-3程式的設計 29
2-4實驗系統架設 31
2-4-1微流體晶片 32
2-4-2剪應力刺激 33
2-4-3正向應力刺激 33
2-4-4程式系統控制 33
2-5細胞繁殖培養 34
2-5-1藥品配製 35
2-5-2細胞繼代 37
2-5-3冷凍細胞與解凍 38
2-6晶片培養細胞流程 39
2-6-1微流體晶片滅菌 39
2-6-2細胞植入晶片 41
2-6-3細胞數量計算 42
第三章結果與討論 43
3-1晶片設計 43
3-2實驗流程 44
3-3剪應力與正向應力參數設定 45
3-3-1剪應力 45
3-3-2正向應力 46
3-3-3剪應力與正向應力實驗 46
3-4檢測正向應力方法 47
3-4-2檢測步驟 47
3-4-3檢測結果 50
3-5檢測剪應力與晶片方法 53
3-5-1實驗步驟與結果 54
3-6細胞受到機械刺激後之結果 55
3-6-1剪應力刺激細胞影響 56
a、施予S1-N1(0.0018 dyne/cm2&0Kpa) 57
b、施予S2-N1(0.044 dyne/cm2&0Kpa) 57
c、施予S3-N1(0.66 dyne/cm2&0Kpa) 57
d、施予S1-N2(0.0018 dyne/cm2&10Kpa) 59
e、施予S2-N2(0.044 dyne/cm2&10Kpa) 59
f、施予S3-N2(0.66 dyne/cm2&10Kpa) 59
g、施予S1-N3(0.0018 dyne/cm2&100Kpa) 60
h、施予S2-N3(0.044 dyne/cm2&100Kpa) 61
i、施予S3-N3(0.66 dyne/cm2&100Kpa) 61
3-6-2正向應力刺激細胞影響 63
第四章結論與未來展望 65
4-1結論 65
4-2未來展望 66
參考文獻 67

參考文獻

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指導教授

曹嘉文

審核日期

2014-12-10

推文