Efficient formation of uniform-sized embryoid bodies using a compartmentalized microchannel device
Efficient formation of uniform-sized embryoid bodies using a compartmentalized microchannel device
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DOI：
10.1039/b618439a
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发表时间：
2007-06-01
期刊：
影响因子：
6.1
通讯作者：
Takayama, Shuichi
中科院分区：
文献类型：
--
作者：
Torisawa, Yu-suke;Chueh, Bor-Han;Takayama, Shuichi
胚状体（EBs）细胞的球形聚集体的形成是胚胎干细胞向其他细胞类型分化的一个必不可少的步骤。适当的形态和胚胎大小对自然受孕胚胎的顺序发育阶段至关重要。同样，调节EBs的大小和形成时间对于控制EB内ES细胞的分化至关重要。然而，现有的生成EBs的方法要么冗长乏味，要么提供大小不一的EBs。在这里，我们描述了一个微流控系统，用于直接同步形成均匀尺寸的电子束，其尺寸可以通过改变微流控装置中微通道的横截面尺寸来控制。该装置由两个微通道组成，由半多孔聚碳酸酯膜隔开，以抵抗细胞粘附。引入上部通道的ES细胞自聚集形成均匀大小的EBs。半多孔膜还允许用来自较低通道的不同试剂对未附着的EBs进行后续处理，而不需要因为膜提供的区隔而冲洗。该方法提供了一种简单而稳健的方法来控制EBs的形成和随后的胚胎干细胞分化，其格式与芯片上的胚胎干细胞处理兼容。
The formation of spherical aggregates of cells called embryoid bodies (EBs) is an indispensable step in many protocols in which embryonic stem (ES) cells are differentiated to other cell types. Appropriate morphology and embryo size are critical for the sequential developmental stages of naturally conceived embryos. Likewise, regulating the size of EBs and the timing of their formation is crucial for controlling the differentiation of ES cells within the EB. Existing methods of formation of EBs, however, are tedious or provide heterogeneously-sized EBs. Here we describe a microfluidic system for straightforward synchronized formation of uniform- sized EBs, the size of which can be controlled by changing the cross-sectional size of rnicrochannels in the microfluidic device. The device consists of two microchannels separated by a semi-porous polycarbonate membrane treated to be resistant to cell adhesion. ES cells introduced into the upper channel self-aggregate to form uniformly-sized EBs. The semi-porous membrane also allows subsequent treatment of the non-attached EBs with different reagents from the lower channel without the need for wash out because of the compartmentalization afforded by the membrane. This method provides a simple yet robust means to control the formation of EBs and the subsequent differentiation of ES cells in a format compatible for ES cell processing on a chip.