藉由整合組織工程及介電泳操控之肝臟細胞晶片於相異培養液濃度環境之研究

On-Chip Patterned Liver Cell Study under Different Concentration of Culture Media

陳鍵瑜(Chien-Yu Chen)

指導教授 ： 劉承賢

國立清華大學/工學院/動力機械工程學系/碩士(2008年)

摘要

隨著微機電產業的蓬勃發展，實驗室晶片技術已變成近期的熱門領域。由於微機電元件符合生物分子尺度，因此在過去數十年間，吸引了許多工程與生物研究者的興趣，投入許多心血與努力，促成跨領域的結合。人工肝臟的建立是近年來在組織工程上一個重要的趨勢，但是希望大量且快速的建立並仿造出肝小葉的圖形，仍有一定的難度 。此外，觀察肝臟細胞在不同濃度的藥物下，活性會如何的變化，也是一個被重視的議題。本論文希望重建肝小葉組織，並研究肝小葉結構在不同濃度藥物下的活性反應行為。 本研究為設計一實驗室晶片，透過晶片上電極的設計，藉由施予適當頻率的交流電壓，使細胞表面被極化成電偶極的基礎下，利用介電泳概念排列細胞，建立肝小葉的圖形。在此過程中，存活率是一個重要的關鍵，因此我們利用FDA/EtBr試劑來完成存活率實驗。在肝小葉結構排列完成後，再經由簡單的微流道設計來提升液體擴散效率，產生一個穩定且連續的藥物濃度梯度，最後流至四個生物反應器，產生不同濃度的藥物，並觀察肝臟細胞活性反應。以上所有功能皆以簡單的製程整合於一晶片上。

關鍵字

微流道 ； 濃度梯度 ； 生物反應器 ； 介電泳 ； 操控 ； 肝小葉

並列摘要

With the development of Micro-Electro-Mechanical Systems (MEMS) industry, the Lab-on-a-chip technology becomes a popular field in recent days. Because the
scale of micro-device matches with cellular molecule,
the interdisciplinary combinations of biological and engineering fields have been attracting a significant
attention in the past few decades. The establishment
of artificial liver is a trend in tissue engineering.
But it is still difficult to build and mimic a hepatic lobule rapidly. Besides, it is an important issue to observe the liver cells activity differences in different concentrations of drug. Rebuilding hepatic lobule and observing the actions of liver cells are the main targets in this thesis.
This research presents a Lab-on-a-chip device to achieve the cell-patterning function. Based on the
polarity difference within cells caused by applying
suitable frequency of the input voltage, we could use
the dielectrophoresis (DEP) to manipulate cells and
form the pattern of the hepatic lobule. During the patterning process, the viability of cells is a key
issue, so we use FDA/EtBr assay to accomplish the
viability tests. After the cell-patterning is finished, stable and continuous concentration gradient of drug is generated by simple microchannel design. Then, different concentrations of culture media are injected into four bioreactors so that the concentrations of culture media
are different in four bioreactors. Finally, we observe
the effect for liver cells by different concentrations
of culture media.

並列關鍵字

Microchannel ； Concentration gradient ； Bioreactor ； Dielectrophoresis (DEP) ； Manipulation ； Hepatic lobule

參考文獻

[71] Jin Jun Chen, Guo Sheng Chen, Nigel J. Bunce,

[9] H. Mohamed, L.D. McCurdy, D.H. Szarowski, S. Duva, J.N.

Kenta Higuchi, Akifumi Kiyota and Ryuichi Ueoka,

[56] Naoya Kobayashi, Teru Okitsu and Noriaki Tanaka,

[72] Lubet R.A., Nims R.W., Mayer R.T., Cameron J.W. and