用於細胞配對與融合之微流體技術發展

Abstract

現今融合細胞，已被廣泛應用於生物醫學相關研究，例如誘導形成幹細胞、癌症免疫療法、生產單株抗體、基因圖譜及組織再生等。在融合過程中，最關鍵的步驟，就是確保細胞配對時能緊密接觸，以提高融合效率。在本論文中，我們提出了一種微流體細胞配對融合晶片，該晶片能透過流體動力學(Hydrodynamics)與微結構抓取與配對細胞，研究中利用鞘流(Sheath flow)與斯托克陷阱(Stokes trap)的方式，進行細胞集中與配對，在鞘流與中心流流速比4：1的情況下，能將細胞集中於20 μm以內。研究中所使用之細胞為NIH-3T3，在細胞密度為2×105 cells/ml時，平均配對時間約為20秒。該晶片能透過流場對細胞施加正向應力，使細胞被擠壓，由原本的圓形擠壓成橢圓形，藉以增加配對時細胞之間的接觸面積，在接觸面積增加的情況下，應可增加細胞融合之效果，並應用於後續細胞融合之研究。

Fusion cells have been extensively used in biomedical research, such as induced stem cell formation, cancer immunotherapy, monoclonal antibodies production, gene mapping, and tissue regeneration. In the process of fusion, the key step is to ensure that cells are in close contact when they are paired, which increase the efficiency of fusion. In this thesis, we propose a microfluidic chip to capture andpair cells by using hydrodynamics and microstructure. In this research, the design of the microfluidic chip is based on sheath flow and Stokes trap. The sheath flow method is used for gathering cells, and the Stokes trap creates a flow field that applies normal stress to increase the contact area of cells when pairing. Under the condition of sheath flow and central flow ratio of 4:1, the cells can be centralized within 20 μm. The cells used in the research are NIH-3T3. The average pairing time is about 20 seconds when the density of the cells is 2×105 cells/ml. The chip can apply positive stress on the cells through the flow field, which squeezed the cells from the original circle to an ellipse so as to increase the contact area between the cells during pairing. When the contact area increases, it should enhance the effect of cell fusion and apply to the subsequent research of cell fusion.