Fan Lei, Guan Zhangyan, Luo Tao, Ren Jifeng, Lam Raymond H W, Sun Dong
Department of Biomedical Engineering, City University of Hong Kong, Hong Kong, China.
Department of Mechanical and Electrical Engineering, Xiamen University, Xiamen, China.
Biomicrofluidics. 2021 Oct 26;15(5):054103. doi: 10.1063/5.0066668. eCollection 2021 Sep.
Single-cell level coculture facilitates the study of cellular interactions for uncovering unknown physiological mechanisms, which are crucial for the development of new therapies for diseases. However, efficient approaches for high-throughput deterministic pairing of single cells and traceable coculture remain lacking. In this study, we report a new microfluidic device, which combines hydrodynamic and recirculation flow captures, to achieve high-throughput and deterministic pairing of single cells in a microwell array for traceable coculture. Compared with the existing techniques, the developed device exhibits advantages with regard to pairing efficiency, throughput, determinacy, and traceability. Through repeating a two-step method, which sequentially captures single cells in a meandering channel and a microwell array, cell number and type can be easily controlled. Double and triple single-cell pairings have been demonstrated with an efficiency of 72.2% and 38.0%, respectively. Cellular engulfment using two breast cell lines is investigated on a developed microfluidic chip as a biological case study, in which the morphological characteristics and the incidence rate are analyzed. This research provides an efficient and reliable alternative for the coculture of single cells on the microfluidic platform for various biomedical applications, such as studying cellular engulfment and tumor sphere formation under single-cell pairing condition.
单细胞水平的共培养有助于研究细胞间相互作用，以揭示未知的生理机制，这对于开发新的疾病治疗方法至关重要。然而，目前仍缺乏用于单细胞高通量确定性配对和可追踪共培养的有效方法。在本研究中，我们报告了一种新型微流控装置，该装置结合了流体动力学和再循环流捕获技术，可在微孔阵列中实现单细胞的高通量确定性配对，用于可追踪的共培养。与现有技术相比，所开发的装置在配对效率、通量、确定性和可追踪性方面具有优势。通过重复两步法，即在蜿蜒通道和微孔阵列中依次捕获单细胞，可以轻松控制细胞数量和类型。已证明双细胞和三细胞单细胞配对的效率分别为72.2%和38.0%。作为一个生物学案例研究，在开发的微流控芯片上研究了使用两种乳腺癌细胞系的细胞吞噬作用，并分析了其形态特征和发生率。本研究为在微流控平台上进行单细胞共培养提供了一种高效可靠的替代方法，可用于各种生物医学应用，如在单细胞配对条件下研究细胞吞噬作用和肿瘤球形成。
