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（hPSC）由于其无限自我更新和分化为几乎任何类型的细胞的独特特性而具有很高的再生治疗潜力。具体地，心肌细胞（CM）现在可以在无异种条件下以高效率从hiPSC衍生，用于心脏修复。然而，在所得细胞群中即使存在少量的非心脏细胞，特别是未分化细胞也是有问题的，因为它们可能在患者中以不可控的方式生长和分化，从而引起畸胎瘤形成的高风险。以前，磁激活细胞分选（MACS）和荧光激活细胞分选（FACS）已用于将未分化的hPSC与靶细胞分离。然而，这些方法昂贵且复杂。在这项研究中，我们挑战性地使用微工程平台去除未分化的hPSC用于衍生的心肌细胞。在此，我们报告了一种具有集成和表面功能化的渔网状结构的微流体装置，用于特定的细胞捕获。在流动衍生表面图案的帮助下，溶液中的细胞被迫穿过渔网状结构，从而实现所选细胞群的高效率和选择性保留。使用加标在培养基或含hiPSC衍生CM的培养基中的hiPSC悬浮液进行器械功能验证。我们发现hiPSC捕获率高达80%，并且在回收的悬浮液中CM群体率显著增加，而不影响细胞活力。

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.