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

人类多能干细胞(HPSCs)具有无限期自我更新和向几乎任何类型的细胞分化的独特特性，因此具有很高的再生治疗潜力。具体地说，心肌细胞(CMS)现在可以在无异种条件下高效地从hPSCs中分化出来，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，在产生的细胞群中也是有问题的，因为它们可能在患者体内以不可控的方式生长和分化，导致形成畸胎瘤的高风险。以前，磁激活细胞分选(MACS)和荧光激活细胞分选(FACS)被用来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，我们的挑战是利用微工程平台去除来源心肌细胞的未分化hPSCs。在这里，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控设备，用于特定的细胞捕获。在流动衍生表面图案的帮助下，溶液中的细胞被迫穿过鱼网状结构，从而导致高效率和选择性地保留选定的细胞群体。在培养液或含有HIPSC来源的CMS的培养液中添加的HiPSCs悬液用于设备功能验证。我们发现，在不影响细胞存活率的情况下，回收的悬液中的HiPSC捕获率高达80%，CM群体比率显着增加。

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.