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

利用微加工技术开发从分化的心肌细胞中去除未分化的人类多能干细胞的系统

基本信息

批准号：
14F04046
负责人：
陳勇
金额：
$ 1.47万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for JSPS Fellows
财政年份：
2014
资助国家：
日本
起止时间：
2014-04-25 至 2016-03-31
项目状态：
已结题

项目摘要

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.

人类多能干细胞（HPSC）由于无限期的自我更新和与任何类型的细胞的独特特性而具有很高的再生疗法潜力。具体而言，现在可以在无XENO条件下以高效的hipsc派生心肌细胞（CMS），朝着心脏修复。然而，在结果的细胞群中，即使存在少量的非心脏细胞，尤其是未分化的细胞也存在问题，因为它们可能在患者中以无法控制的方式生长和区分，从而导致畸胎瘤形成的高风险。以前，磁性细胞分选（MAC）和荧光激活的细胞分选（FACS）已用于将未分化的HPSC与靶细胞分开。但是这些方法很昂贵且复杂。在这项研究中，我们挑战要使用微工程平台去除未分化的HPSC来衍生的心肌细胞。本文中，我们报告了一种微流体装置，具有集成和表面功能化的渔网样结构，用于特定的细胞捕获。借助流导向表面模式，溶液中的细胞被迫越过类似鱼网的结构，从而高效效率并选择性地保留所选的细胞群。在培养基或含有培养基的HIPSC衍生CMS中尖刺的HIPSC悬浮液用于设备功能验证。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回收悬浮液中CM人口率显着增加。