Three-dimensional spheroid-forming lab-on-a-chip using micro-rotational flow
Three-dimensional spheroid-forming lab-on-a-chip using micro-rotational flow
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DOI：
10.1016/j.snb.2009.11.061
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发表时间：
2010-05-18
影响因子：
8.4
通讯作者：
Miki, Norihisa
中科院分区：
文献类型：
--
作者：
Ota, Hiroki;Yamamoto, Ryosuke;Miki, Norihisa
我们提出了一个球体形成的芯片实验室，使用微旋转流来控制三维肝细胞球体的大小。所开发的装置包括由软光刻产生的聚二甲基硅氧烷制成的微室和由储液器、阻尼器、室、粉碎器通道和蠕动泵组成的灌注系统，以不断地循环含有人肝细胞肝癌细胞的细胞培养基。细胞被吸引到腔室的中心，在那里它们聚集并形成球状体。通过改变细胞密度和流动条件，可以控制由该装置产生的球状体的大小。因此，可以在不改变腔室设计的情况下形成各种尺寸的球状体。所开发的灌注系统允许培养物通过抑制微旋转流的突然变化而储存很长时间;这种能力通过进行活/死测定来验证。成功地产生了平均直径在130-430 μ m范围内，标准偏差小于17.2%的肝球状体。发现它们的大小取决于腔室直径和细胞密度。平均直径在150 ~ 200 μ m的球状体的标准偏差低至16.0%。显微粒子图像测速显示了一个球状体形成区域的室。它还能够估计形成球体的细胞数量。皇冠版权所有（C）2010由爱思唯尔B. V.出版保留所有权利。
We propose a spheroid-forming lab-on-a-chip that uses micro-rotational flow to control the size of three-dimensional hepatocyte spheroids. The developed device consists of a microchamber made of polydimethylsiloxane produced by soft lithography and a perfusion system consisting of a reservoir, a dampener, a chamber, a shredder channel, and a peristaltic pump to constantly circulate cell culture medium containing human hepatocellular liver carcinoma cells. Cells are attracted to the center of the chamber, where they aggregate and form spheroids. The size of the spheroids produced by this device can be controlled by varying the cell density and the flow conditions. Thus, spheroids of various sizes can be formed without altering the chamber design. The developed perfusion system permits cultures to be stored for a long time by suppressing sudden changes in the micro-rotation flow; this ability was verified by performing a live/dead assay. Hepatic spheroids with average diameters in the range 130-430 mu m and with a standard deviation of less than 17.2% were successfully produced. Their sizes were found to depend on the chamber diameter and the cell density. The standard deviation of spheroids with average diameters between 150 and 200 mu m was as low as 16.0%. Micro particle image velocimetry revealed a spheroid-forming region in the chamber. It also enabled the number of cells that form a spheroid to be estimated. Crown Copyright (C) 2010 Published by Elsevier B.V. All rights reserved.