這個研究中所研發的癌症免疫療法微環境實驗室晶片，結合有機光導電材料的氧化鈦酞菁TiOPc來製作光電鑷夾並建立以光圖形來操控免疫細胞及癌症細胞並克服微流體中的問題，讓細胞跟細胞之間可以緊密接觸，進而提供免疫細胞和癌症細胞之間的相互作用研究並動態分析自然殺手細胞活性。在微流道中，容易有流體停滯死區問題並不容易動態觀察細胞間相互作用，所以，藉由創造出穩定和靜態的培養微環境，可以減少流場引發的剪應力效應並增強自然殺手細胞的活性而達到交互作用。另外，結合使用無光罩曝光機，將聚乙二醇二丙烯酸酯水凝膠（PEG-DA）光聚合固化，製做四葉草形狀（FLCS）微孔環境，開發運用光電鑷夾操控單一細胞至微孔環境裡面，以進行自然殺手細胞與標靶細胞交互作用，同時解決微流體中停滯死區問題。本博士論文研究結合多種前瞻技術的研發，解決及重建了一個體外人體免疫系統微環境，用於研究動態免疫反應。當自然殺手細胞在微環境實驗室晶片中與標靶細胞接觸時，這個設計能即時觀察到標靶細胞表現出凋亡特徵（即在2小時內細胞收縮和起泡，然後在3小時內死亡）。與傳統生醫實驗室常規分析相比，這個微環境實驗室晶片表現出更高的自然殺手細胞活性，在自然殺手細胞跟標靶細胞比例皆為一比一時，自然殺手細胞殺掉標靶細胞的比例比傳統的高出7.7% ; 在自然殺手細胞跟標靶細胞比例為十比一時，自然殺手細胞殺掉標靶細胞的比例比傳統的高出5.1%。同時，本博士論文研究也展示了我們開發的癌症免疫療法微環境實驗室晶片可以增強自然殺手細胞活性的潛力，並動態研究免疫細胞與癌細胞間的免疫相互作用。

A cancer immunotherapy μ-environment LabChip, equipped with titanium oxide phthalocyanine (TiOPc)-based optoelectronic tweezers (OET) to achieve direct cell–cell contact, was developed in this Ph.D. study and could be applied to study the interaction between immune cells and other cells. One of the applications as demonstrated for the real-time analysis of nature killer (NK) cells' behavior. In microfluidic devices, it is difficult to simultaneously solve the dead zone problems and observe dynamic cell-cell interactions. A stable and static culture μ-environment LabChip was focused and developed in this Ph.D. research to enhance NK cell activities to explore cancer immunotherapy studies. In addition, OET was developed and used to solve the dead zone problems by manipulating a single cell into four-leaf-clover-shaped (FLCS) microwells made of light-activated hydrogel, Poly(ethylene glycol) diacrylate (PEG-DA), through optofluidic maskless lithography, to enhance direct cell-cell contact. This design reconstructed an in vitro human immune system for the studies of dynamic immunological response. When the NK cells came into contact with the target cells in the μ-environment LabChip, the target cells showed apoptotic characteristics (i.e. cell shrinkage and blebbing within 2 h and then die within 3 h). The NK cell activity higher than 7.7%, compared with the NK cell: target (1:1) under the conventional analysis, was demonstrated via the μ-environment LabChip developed in this Ph.D. study. An innovative cancer immunotherapy μ-environment LabChip has been developed to provide a stable and static environment for cell–cell interaction studies. Furthermore, this μ-environment LabChip shows the potential to enhance NK cell activity and to study immunological interactions between immune cells and cancer cells dynamically.

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