人類肝臟細胞在高分子基材上之行為表現及細胞電永在組織工程上的應用

The study pf behaviors of hepatocyte on different biomaterials and the application of macroelectrophoresis on tissue engineering

陸瑞男(Jui-Nan Lu)

指導教授 ： 楊台鴻

國立臺灣大學/工學院/醫學工程學研究所/博士(2011年)

https://doi.org/10.6342/NTU.2011.00067

摘要

在此研究中，我們利用高分子基材培養冷凍四年的人類肝臟細胞，結果顯示肝臟細胞在冷凍四年後在聚乙烯醇上仍然可以保有肝臟細胞的功能及活性，因此我們認為聚乙烯醇是一個未來能夠應用在培養冷凍肝臟細胞的有潛力之高分基材。在第二部分我們以同時含有幾丁聚醣及組織培養的聚苯乙烯之複合材料培養環境去培養人類肝臟的細胞株HepG2，從實驗中我們可以知道決定一個細胞的行為表現主要的因素是來自於細胞基底下面的基材，景館一個環境中還有兩種行為表現不同的細胞存在，彼此並不會相互影響而改變細胞在基材上的特性。在第三部分，我們利用胰島素的添加去改變細胞在幾丁聚醣上的行為表現，從實驗中我們知道利用胰島素的添加可以明顯的提升肝臟細胞在幾丁聚醣上的貼付能力與增生的能力，此外我們同樣的利用抑制PI-3 kinase路徑的藥劑來終止胰島素對細胞行為的影響。在最後的部分，我們利用細胞電泳來偵測神經幹細胞初期的分化。此外我們發現神經幹細胞初期的分化會伴隨著表面電位的提升。

關鍵字

人類肝細胞 ； 冷凍保存 ； HepG2細胞株 ； 幾丁聚醣 ； 細胞電泳

並列摘要

The purpose of the first part was to evaluate the behaviors of long-term (more than 4 years) cryopreserved human hepatocytes on biomaterials. Human hepatocytes could be cryopreserved for more than 4 years without losing liver-specific function and PVA was proposed to serve as an appropriate and promising substrate for the use in culturing long-term cryopreserved hepatocytes by maintaining high cell attachment and of high level of liver-specific function. In the second part, cells were cultured on a multi-biomaterials substrate (C/T substrate) both composed of chitosan and TCPS (tissue culture polystyrene) for 8 days. Cells expressed the behaviors based on the substrate under the cells, and none of cell was found that migrated from one substrate to the other substrate. And in the third part, insulin was added into the CT substrate to control the cell behaviors on the chitosan. Insulin greatly promted cell adhesion and enhanced the proliferation on chitosan. The insulin induced cellular behaviors can be removed by blocking the PI-3 kinase pathway, and cell behaviors restored to the chitosan-induced behaviors. And in the final part, macroelectrophoresis was used to detected the differentiation of neural stem/progenitor cell (NSPC). We found that the surface charge of NSPC was up-regulated during the early period of differentialtion.

並列關鍵字

hepatocyte ； cryopreservation ； HepG2 ； chitosan ； macroelectrophoresis

參考文獻

Background

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