微流體細胞分注裝置應用於高通量藥物篩選之研發

Abstract

在傳統藥物篩選流程中，人工分注細胞的過程不僅耗時，同時也限制實驗通量的提升。為此，本研究開發了一應用於高通量藥物篩選的微流體細胞分注裝置，可實現快速且自動化地將細胞分注至實驗孔盤中。本裝置設計基於微流道中流體所呈現的層流特性，以 Hagen-Poiseuille 方程式進行幾何設計以達下游流道等效流阻，使細胞懸浮液於注射幫浦所產生之壓力驅動下，透過流道結構引導至各出口。裝置同時引入被動式擾動器，提升出口細胞的均一性。在裝置性能的驗證中，研究使用 10 μm 之聚苯乙烯微粒作為細胞的模擬模型，初步確認裝置均勻分流性能。後續使用人類結腸腫瘤癌細胞株 (HCT-116) 進行細胞的分流測試，並透過 AlamarBlue 試劑進行細胞活性的測試。最後以化療藥物 5-FU (5-Fluorouracil) 對 HCT-116 進行細胞毒性的測試，以驗證本裝置於高通量藥物篩選流程中的應用可行性。實驗結果顯示，此微流體裝置可於 2 分鐘內完成 96 孔盤的分注，且輸出之總粒子數量與目標值誤差控制於 7.933% 以內。後續細胞分流實驗亦顯示輸出至孔盤之細胞數量具有高再現性，且細胞經裝置分注後可正常貼附，未見變形與污染現象，顯示裝置具備良好的操作安全性與生物相容性。最終，細胞毒性測試展示經裝置分注之細胞可保有對藥物刺激的反應能力，證實裝置適用於高通量藥物篩選場景。本研究開發之微流體裝置可在短時間內完成細胞分注，預期其可在藥物篩選領域為中小型研究單位與臨床場域提供一低成本、高效率的自動化平臺。

Manual cell dispensing in conventional drug screening workflows is not only labor-intensive, but also limits overall experimental throughput. To address this bottleneck, this study develops a microfluidic cell dispensing device for high-throughput drug screening, enabling rapid and automated cell dispensing into well plates. To ensure uniform flow distribution across outlets, the device leverages fluid's laminar flow characteristics within microchannels, where downstream channel geometries are designed according to the Hagen-Poiseuille equation to achieve equivalent hydraulic resistance. Driven by a single syringe pump, cell suspension is guided through the channel network to multiple outlets. Furthermore, integrated passive perturbation structures enhance the uniformity of the dispensed cells at the outlets. The device's performance is initially validated using 10 µm polystyrene beads as a cell mimic, confirming its capability for uniform distribution. Subsequently, human colon carcinoma cell line (HCT-116) is used to evaluate cell aliquoting performance, with cell viability assessed via the AlamarBlue assay. Ultimately, chemotherapeutic drug 5-Fluorouracil (5-FU) is used to perform a cytotoxicity test on the dispensed HCT-116 cells, validating the device's applicability in a high-throughput drug screening workflow. Results demonstrate that the microfluidic device can complete dispensing a 96 well plate within 2 minutes, with the deviation of the total particle count at each outlet controlled within 7.933% of the target value. The device exhibits good biocompatibility, operational consistency, and high reproducibility in dispensing viable cells, as cells adhere properly with no observable deformation or contamination. Lastly, cytotoxicity test shows cells dispensed by the device retained their ability to respond to drug stimuli. This work presents an efficient solution for cell dispensing in high-throughput drug screening, expects to offer a cost-effective platform for biomedical research and clinical settings in drug screening.