研究生: |
朱巧文 Chu,Chiao Wen |
---|---|
論文名稱: |
以纖維狀水膠三維培養腎足細胞 Cultivating Podocytes in fibre-shaped hydrogel |
指導教授: |
曾繁根
Tseng,Fan Gang |
口試委員: |
許翔皓
鄭兆珉 |
學位類別: |
碩士 Master |
系所名稱: |
原子科學院 - 工程與系統科學系 Department of Engineering and System Science |
論文出版年: | 2015 |
畢業學年度: | 103 |
語文別: | 中文 |
論文頁數: | 61 |
中文關鍵詞: | 腎足細胞 、微管 、海藻膠 、細胞支架 |
外文關鍵詞: | podocyte, microfiber, alginate, scaffold |
相關次數: | 點閱:2 下載:0 |
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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.
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