光誘導細胞融合之微流體晶片

Optically-induced Cell Fusion on a Microfluidic Chip

楊博夫(Yang, Po Fu)

指導教授 ： 李國賓

國立清華大學/工學院/動力機械工程學系/碩士(2015年)

摘要

細胞融合等技術目前被廣泛應用於基因重組、製造單株抗體、癌症免疫療法以及組織再生等各項生物應用，而現今細胞融合可利用生物、化學與物理方法達成。但是在傳統大型系統上，由於細胞間的不穩定接觸與隨機配對上的限制使得效率與產量上難以提升。因此，有效的增加細胞接觸與配對是相當關鍵之因素來提升細胞融合的效率與產量。本研究提出一新方法，透過整合微結構與光誘導技術於微流體系統來穩定細胞間的接觸與配對以達到精確、高效率、高產量的細胞融合。而光誘導技術是將設計好之圖形利用投影機將其投射在一鍍有非晶矽薄膜的氧化銦錫玻璃上，被照射之非金矽薄膜區域的導電度會提升數個量級，並同時在上下兩側之氧化銦錫玻璃上導入交流電場，如此一來在受光照射之非晶矽薄膜上之會產生局部之加強電場，進而形成”虛擬電極”，藉此來取代傳統系統上的固定微電極，使得能輕易且彈性的操縱電極之數量、大小、位置等參數來提升效率與產量，此虛擬電極已經成功地應用於細胞分離、細胞裂解、細胞電穿孔等應用。故本研究創造一新方法，先利用微流道上之微結構所產生之流場將細胞依序抓取且配對，並達到57%的配對率，再透過整合光誘導技術之虛擬電極使細胞融合效率達到87%。因此，光誘導細胞融合系統是一項可以進行高效率與高產量之細胞融合的先進技術。

關鍵字

細胞配對 ； 細胞融合 ； 光誘導細胞融合 ； 微流體系統

並列摘要

Cell fusion is a critical course for all sort of biomedical applications including cell reprogramming, hybridoma formation, cancer immunotherapy, and tissue regeneration. It can be realized by using biological, chemical, or physical methods. However, efficiency and yields are limited by unstable cell contact and random cell pairings in traditional methods. Hence, improving cell contact and cell pairing are the two key factors to enhance efficiency and yields of cell fusion. This study therefore reported a new approach called optically-induced cell fusion (OICF) which integrates cell-pairing microstructures and optically-induced, localized electrical field to achieve precise cell fusion with high yields and high efficiency. By projecting light patterns on a photoconductive film (hydrogen-rich amorphous silicon, a-Si: H) coated on an indium-tin-oxide (ITO) glass while an alternating-current (AC) electric field was applied on the top and bottom ITO glasses, “virtual” electrodes would be constructed accordingly. In fact, this method could be used on several biomedical applications, including cell manipulation, cell separation, cell lysis and electroporation. Therefore, a locally enhanced electric field would be induced and the pairing cells could be precisely fused by the virtual electrodes. In this study, 57% cell paring rate and 87% fusion efficiency were achieved. Therefore, OICF is a promising method to succeed in cell fusion with high efficiency and high yields.

並列關鍵字

cell pairing ； cell fusion ； optically-induced cell fusion ； microfluidics

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