微細加工技術を用いた分化心筋細胞からの未分化ヒト多能性幹細胞除去システムの開発

人多能干细胞（hPSCs）具有无限自我更新和分化为几乎任何类型细胞的独特特性，在再生治疗中具有很高的潜力。具体来说，现在可以在无xeno条件下高效地从hipsc中提取心肌细胞（CMs）用于心脏修复。然而，即使是少量的非心脏细胞，特别是未分化的细胞，在产生的细胞群中存在问题，因为它们可能在患者体内以不可控的方式生长和分化，导致畸胎瘤形成的高风险。此前，磁激活细胞分选（MACS）和荧光激活细胞分选（FACS）已被用于从靶细胞中分离未分化的hPSCs。然而，这些方法既昂贵又复杂。在这项研究中，我们正在挑战使用微工程平台为衍生心肌细胞去除未分化的hPSCs。在此，我们报告了一种具有集成和表面功能化鱼网状结构的微流体装置，用于特定的细胞捕获。在流动衍生表面模式的帮助下，溶液中的细胞被迫穿过鱼网状结构，从而产生高效率和选择性保留选定的细胞群。将hiPSC悬液加在培养基中或hiPSC衍生的含有培养基的CMs中用于设备功能验证。我们发现hiPSC的捕获率高达80%，并且在不影响细胞活力的情况下，恢复的悬浮液中CM群体率显着增加。

Human pluripotent stem cells (hPSCs) hold high potential for regenerative therapies due to their unique properties of indefinite self-renewal and differentiation to almost any type of cells. Specifically, cardiomyocytes (CMs) can now be derived from hiPSCs at high efficiency under xeno-free condition toward heart repairing. However, the presence of even small numbers of non-cardiac cells, particularly the undifferentiated cells, in the resulted cell population, is problematic as they may grow and differentiate in an uncontrollable manner in the patient, giving rise to high risk of teratoma formation. Previously, magnetic-activated cell sorting (MACS) and Fluorescence-activated cell sorting (FACS) have been used to separate undifferentiated hPSCs from target cells. However those methods are expensive and complicated. In this study, we are challenging to remove undifferentiated hPSCs for derived cardiomyocytes using micro-engineered platform. Herein we report a microfluidic device with integrated and surface functionalised fishnet-like structures for specific cell capture. With the help of a flow derivation surface pattern, cells in solution are forced to cross the fishnet-like structure, resulting in high efficiency and selective retention of a chosen cell population. A suspension of hiPSCs spiked in culture medium or hiPSC derived CMs containing medium was used for devices function validation. We found a hiPSC capture rate as high as 80% and a remarkable increase of the CM population rate in the recovered suspension without affecting the cell viability.