腎臟足細胞（Podocyte）是位於腎小球毛細血管基底膜外層終端分化的細胞，其足突間的裂孔膜（Split Mernbrane）是腎小球過濾的最後一道屏障，當足細胞受傷時，足細胞的蛋白濾過屏障完整性會被破壞，造成大量蛋白尿的產生。隨著對足細胞生物學研究的深入，尤其是在發現足細胞表達的一些特異性蛋白分子後，足細胞已被認為是參與各種原發性或繼發性腎小球疾病進展的關鍵細胞。
本研究希望開發出一種具彈性及透析能力之管狀生物支架，以釐清腎小球損傷與腎臟衰竭之關連性及設計具彈性伸縮之微米結構平台產生動態機械刺激腎足細胞之研究。我們利用高生物相容性的明膠與海藻膠作為細胞的立體支架，以有較好機械強度的海藻膠作為外殼，並將細胞混合明膠與膠原蛋白作為芯層，以層流方式與海藻膠同時注入氯化鈣水溶液中，利用海藻膠與鈣離子結合立即固化之特性將細胞培養於管中，同時明膠會在培養箱的環境中(33℃)變回液態，而使此管狀結構變為中空管狀之生物支架。目前已能夠成功將腎足細胞培養於纖維狀支架中，並且看到其生長，雖然還未找到最適合腎足細胞生長的條件，但能預期細胞在適當的環境下能夠生長成我們所希望之腎小球微血管狀，並未來本研究裝置亦可提供配合臨床藥物研究，設計治療腎臟病變所需之藥物。

The podocyte is a highly specialized cell which is responsible for the glomerular permselectivity of proteins in the kidney. Podocyte foot processes comprise a highly branched interdigitating network with foot processes of neighboring podocytes connected by the slit membrane (SM), which is a multi-protein complex similar to adheren junctions and covers filtration slits, thereby establishing the final barrier to urinary protein loss.
This study desired to develop a flexible and dialysis of the tubular scaffold ability to clarify the connection between glomerular injury and kidney failure.
We use high biocompatibility of gelatin and sodium alginate as a three-dimensional scaffold of cells. In order to have better mechanical strength to contain the fiber-shape, we use sodium alginate as a shell, and the cells were mixed with collagen and gelatin as a core layer. These two solutions were injected into calcium chloride aqueous simultaneously and the flow rates are maintained in the laminar flow. By the properties of sodium alginate, calcium-binding immediately, the cells were cultured in the tube. While the gelatin in the incubator environment (33℃) back into a liquid, thereby enabling the tubular structure into a hollow tubular scaffolds. We have been able to successfully cultured in the fibrous podocytes, and to see their growth, although not yet find the most suitable conditions for the growth of podocytes, but the cells can be expected under appropriate circumstances we can grow into the desired glomerular capillary-like.

總目錄
摘要 II
ABSTRACT III
圖目錄 5
第一章 緒論 9
1.1 研究背景 9
1.2 研究動機 10
1.3 研究目的 11
第二章 文獻回顧 12
2.1 終末期腎病變 12
2.2影響腎足細胞過濾因素 14
2.3壓力對腎足細胞之影響 16
2.4水膠三維空間培養 19
2.5製造管狀支架系統 24
第三章 實驗步驟、設備與材料 28
3.1 細胞微環境操作 28
3.1.1 細胞培養基本技術 28
3.2 多層水膠製程 29
3.2.1 多層水膠製作流程 29
3.3水膠原料與配製 31
3.3.1 GelMa原料配製 31
3.3.2水膠與光起始劑配製溶液 31
3.3.3 甲殼素原料 32
3.3.4 甲殼素薄膜 32
3.3.5明膠原料配製 33
3.3.6海藻膠配製 33
3.4 細胞生長曲線 34
3.4.1 添加鈣離子對腎足細胞之影響 34
3.4.2 添加膠原蛋白於含鈣離子培養液對細胞之影響 34
3.5 水膠中培養細胞 35
3.5.1 二維培養細胞於水膠中生長 35
3.5.2 培養細胞於管狀 35
3.6 實驗儀器與系統 36
第四章 實驗結果與討論 37
4.1 多層結構培養細胞 37
4.1.1 各層結構圖及其作用 37
4.2 水膠三維空間培養 40
4.2.1 血液透析模培養細胞 42
4.3 明膠與海藻膠固化測試 42
4.3.1 明膠 42
4.3.2 海藻膠 44
4.4 流速測試 45
4.4.1 同軸流體 45
4.5 三維培養細胞 47
4.5.1 添加鈣離子對腎足細胞之影響 47
4.5.2 培養腎足細胞於纖維支架 49
4.5.2 培養HeLa於纖維支架 51
4.5.3 改善殼層條件培養細胞 52
第五章 結論 53
5.1 多層結構培養內皮細胞 53
5.2 二維空間培養 53
5.3 纖維支架成形 54
5.4三維水膠空間培養 55
第六章 未來工作 56
參考文獻 58

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