Li Jiyu, Liu Ya, Ren Jifeng, Tay Benjamin Zikai, Luo Tao, Fan Lei, Sun Dong, Luo Guannan, Lau Denvid, Lam Raymond H W
Department of Biomedical Engineering, City University of Hong Kong, Hong Kong.
School of Mechanical and Aerospace Engineering, Nanyang Technological University, Singapore.
Lab Chip. 2020 Mar 17;20(6):1072-1082. doi: 10.1039/d0lc00078g.
Cell isolation from blood is an important process for diagnosing immune diseases. There are still demands for a user-friendly approach to achieve high cell extraction efficiency and purity of a target immune cell subtype for more promising diagnosis and monitoring. For selective immune cell isolation, we developed a microstructured device, which consists of antibody-coated micropillars and micro-sieve arrays, for isolating a target immune cell subtype from bovine blood samples. The focusing micropillars can guide immune cells flowing to the subsequent micro-sieves based on deterministic lateral shifts of the cells. The arrangement of these microstructures is characterized and configured for the maximal cell capture rate. Surface modification with a selected antibody offers selective cell capture in the micro-sieves based on the antigen-antibody reaction. We prepare a cell mixture of human CD14-expressing leukemia cells (THP-1) and epithelial cells (MDA-MB-231) in diluted blood to characterize the cell isolation operation, with a selective cell isolation yield of >80%, cell purity of ∼100% and cell viability of >93%. Together, this microstructured device strategy can achieve high-yield selective isolation of immune cells from blood samples and support downstream genetic and biochemical cell analyses, contributing to the medical diagnosis of a broad range of immune diseases.
从血液中分离细胞是诊断免疫疾病的重要过程。对于更有前景的诊断和监测而言，仍然需要一种用户友好的方法来实现高细胞提取效率以及目标免疫细胞亚型的纯度。为了进行选择性免疫细胞分离，我们开发了一种微结构装置，它由抗体包被的微柱和微筛阵列组成，用于从牛血样本中分离目标免疫细胞亚型。聚焦微柱可基于细胞的确定性侧向位移引导免疫细胞流向后续的微筛。这些微结构的排列经过表征和配置，以实现最大的细胞捕获率。用选定抗体进行表面修饰可基于抗原 - 抗体反应在微筛中进行选择性细胞捕获。我们在稀释血液中制备了表达人CD14的白血病细胞（THP - 1）和上皮细胞（MDA - MB - 231）的细胞混合物，以表征细胞分离操作，选择性细胞分离产率>80%，细胞纯度约为100%，细胞活力>93%。总之，这种微结构装置策略可以实现从血液样本中高产率地选择性分离免疫细胞，并支持下游的细胞遗传和生化分析，有助于对多种免疫疾病进行医学诊断。
