Department of Nanomedicine, Houston Methodist Research Institute, Houston, TX, 77030, USA.
Department of Cell Biology and Medical Genetics, Nanjing University of Chinese Medicine, Nanjing, 210023, P. R. China.
Proteomics. 2020 Jul;20(13):e1900223. doi: 10.1002/pmic.201900223. Epub 2020 Jan 3.
Hematopoietic stem/progenitor cell (HSPC) mobilization from the bone marrow to the bloodstream is a required step for blood cell renewal, and HSPC motility is a clinically relevant standard for peripheral blood stem cell transplantation. Individual HSPCs exhibit considerable heterogeneity in motility behaviors, which are subject to complex intrinsic and extrinsic regulatory mechanisms. Motility-based cell sorting is then demanded to fulfill the study of such mechanism complexity. However, due to the HSPC heterogeneity and difficulty in monitoring cell motility, such a platform is still not available. With the recent development of microfluidics technology, motility-based monitoring, sorting, collecting, and analysis of HSPC behaviors are highly possible and achievable if fluid channels and structures are correctly engineered. Here, a new design of microfluidic arrays for single-cell trapping is presented, enabling high-throughput analysis of individual HSPC motility and behavior. Using these arrays, it is observed that HSPC motility is positively correlated with CD34 asymmetric inheritance and cell differentiation. Transcriptomic analysis of HSPCs sorted according to motility reveals changes in expression of genes associated with the regulation of stem-cell maintenance. Ultimately, this novel, physical cell-sorting system can facilitate the screening of HSPC mobilization compounds and the analysis of signals driving HSPC fate decisions.
造血干细胞/祖细胞（HSPC）从骨髓动员到血液是血细胞更新所必需的步骤，而 HSPC 的迁移能力是外周血干细胞移植的一个临床相关标准。单个 HSPC 在迁移行为上表现出相当大的异质性，这受到复杂的内在和外在调节机制的影响。因此，需要基于迁移的细胞分选来研究这种机制的复杂性。然而，由于 HSPC 的异质性以及监测细胞迁移的困难，这样的平台仍然不可用。随着微流控技术的最新发展，如果正确设计流体通道和结构，基于迁移的 HSPC 行为的监测、分选、收集和分析是非常可能和可行的。在这里，提出了一种用于单细胞捕获的微流控阵列的新设计，能够实现对单个 HSPC 迁移和行为的高通量分析。使用这些阵列，观察到 HSPC 的迁移能力与 CD34 不对称遗传和细胞分化呈正相关。根据迁移能力分选的 HSPC 的转录组分析揭示了与干细胞维持调控相关的基因表达变化。最终，这种新颖的物理细胞分选系统可以促进 HSPC 动员化合物的筛选以及驱动 HSPC 命运决定的信号分析。
