On-chip platelet production using three dimensional microchannel

H. Kumon, S. Sakuma, S. Nakamura, K. Eto, F. Arai
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Abstract

We succeeded in on-chip platelet production using a bioreactor with a curve-shaped 3D microchannel. In order to produce platelets from megakaryocytes (MKs) in a microfluidic chip, it is required to trap MKs and to apply fluid force to it. Since MKs have a relatively big distribution in their size, it is difficult to effectively trap MKs by using conventional microfluidic chips having uniformly patterned pillars with discrete pitch size. Thus, we proposed a curve-shaped 3D microchannel whose height gradually decreases along the flow pass to trap MKs of various size. We fabricated the curve-shaped 3D microchannel by using grey-scale lithography and deep reactive ion etching (DRIE) techniques. Since our microfluidic chip was packaged by a glass substrate, we can observe the processes of platelet production with a time-resolved technique. Through the experiments of on-chip platelet production using MKs induced from human induced pluripotent stem cells (hiPSCs), we successfully trapped the MKs of various size corresponding to the channel height. The trapped MKs were exposed to fluid force in the microchannel, and resulted in producing platelets.
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片上血小板生产利用三维微通道
我们成功地在片上血小板生产使用生物反应器与曲线形状的三维微通道。为了在微流控芯片中从巨核细胞(mk)产生血小板,需要捕获巨核细胞并对其施加流体力。由于mk在尺寸上的分布比较大,传统的微流控芯片很难有效地捕获mk,这些微流控芯片具有均匀的图案柱和离散的节距尺寸。因此,我们提出了一个曲线形状的三维微通道,其高度沿流动通道逐渐降低,以捕获不同尺寸的mk。采用灰度光刻和深度反应离子刻蚀(DRIE)技术制备曲线形状的三维微通道。由于我们的微流控芯片是用玻璃基板封装的,我们可以用时间分辨技术观察血小板的产生过程。通过利用人诱导多能干细胞(hiPSCs)诱导的mk在芯片上生产血小板的实验,我们成功捕获了与通道高度相对应的不同大小的mk。被捕获的mk暴露在微通道中的流体力下,导致血小板的产生。
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