以微機械技術製造圖形式細胞 應用於活體神經訊號量測之研究

Abstract

人為操控神經細胞生長與再生之技術，對於基礎生物學、神經學和組織學研究，一直扮演著很重要的關鍵角色。近十年來，許多生物技術的發展，像是微沾印(μCP)和毛細管微模化(MIMIC)等，皆將焦點著重於製做人工微蛋白質圖形上，並藉由此人工微蛋白質圖形，以誘導(操控)神經細胞生長至所需的位置。 在本論文中，我們提出了一個新式的人工蛋白質圖形產生技術：上開蓋式微流道(Open-Top SU-8 Microfluidic Channel)，藉由此上開蓋式微流道所產生微蛋白質圖形，可引導、束縛神經細胞之生長方向及位置。此種上開蓋式微流道只需少量的蛋白質，約0.5μl，即可填滿兩個蛋白質填充槽和六個微流道，產生良好的微蛋白質圖形。 在神經的培養上，我們選擇了金魚視網膜神經細胞(RGCs)。並成功地培植視網膜神經於上開蓋式微流道內生長，證實「上開蓋式微流道技術」應用於神經操控之可行性。 除此之外，微機械製造的上開蓋式微流道更展現出另一特點：整合度高。本論文中所提的上開蓋式微流道可於製程中整合微電子陣列裝置，此可解決微蛋白質圖形轉移印至微電子裝置，並提供更廣泛的研究於於基礎生物學、神經學和組織學上。

Manipulating neuron outgrowth and regeneration is important and attractive in the research fields of biomedical, neurological and tissue engineering. Over the last ten years, many bio-techniques such as μCP and MIMIC were developed on generating artificial micropatterns of extracellular matrix proteins (ECM) for guiding neuron to designed positions. In this study, we propose a new scheme, i.e. open-top SU-8 microfluidic channel, to generate micropatterns of ECM proteins to guide and control axons outgrowth in preferred directions and at designed positions by confinement in a microfluidic system. The open-top SU-8 microfluidic channel requires only a small volume (~0.5μl) of laminin solution to fill the two tanks and microchannel. The culturing of retinal ganglion cells (RGCs) was performed using injured goldfish. The experimental results verify that the open-top SU-8 microfluidic channel is not only workable but also effective for neural guidance. Besides, the micromachined open-top SU-8 microfluidic channel shows good integration ability of microelectronic device of microelectrode array (MEA) into the microfluidic channels, which solve the problem of transferring micro protein patterns on microelectronic device.