Microfluidic hydrodynamic cellular patterning for systematic formation of co-culture spheroids.

Microfluidic hydrodynamic cellular patterning for systematic formation of co-culture spheroids.
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DOI:
10.1039/b915965g
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发表时间:
2009-12
期刊:
Integrative biology : quantitative biosciences from nano to macro
影响因子:
--
通讯作者:
Takayama S
Takayama S
中科院分区:
其他
文献类型:
--
作者:
Torisawa YS;Mosadegh B;Luker GD;Morell M;O'Shea KS;Takayama S

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本文描述了一种微流控方法,以形成各种几何形状和组成的共培养球体,以操纵细胞-细胞相互作用动力学。细胞图案化在双层微流体装置中进行,该装置夹有半多孔膜,使得从顶部通道通过膜到底部通道发生流动。通过调节底部通道的几何特征使得任意细胞排列成为可能,使得当培养基排出时,流动流体动力学地将细胞聚集(聚集)到仅在底部通道的区域上的膜上。此外,当顶部通道具有多个入口时,细胞可以接种在相邻的层流中,从而允许不同的细胞类型以良好限定的空间布置同时图案化。有趣的是,某些细胞类型的初始细胞定位可以导致两个并列的非同心的“Janus”球体,而不是均匀的混合物或分层的壳结构。因此,细胞在聚集之前的初始位置可以影响共培养球体内的最终构型。当Janus球状体由小鼠胚胎干(mES)细胞和肝细胞构建时,mES细胞以由肝细胞位置决定的空间上不同的模式分化。这与当通过随机混合两种细胞类型的常规方法形成共培养球状体时观察到的均匀mES分化形成对比。这种细胞图案化方法为理解和操纵3D中不同细胞类型之间的相互作用开辟了新的可能性。
This paper describes a microfluidic method to form co-culture spheroids of various geometries and compositions in order to manipulate cell–cell interaction dynamics. The cellular patterning is performed in a two-layered microfluidic device that sandwiches a semi-porous membrane so that flow occurs from the top channel through the membrane to the bottom channel. Arbitrary cellular arrangements are enabled by regulating the geometric features of the bottom channel so that as culture media drains, the flow hydrodynamically focuses (aggregates) cells onto the membrane only over the regions of the bottom channel. Furthermore, when the top channel has multiple inlets, cells can be seeded in adjacent laminar streams, allowing different cell types to be patterned simultaneously in well defined spatial arrangements. Interestingly, the initial cell positioning of certain cell types can result in two juxtaposed non-concentric “Janus” spheroids, rather than homogeneous mixtures or layered shell structures. Therefore, the initial position of cells prior to aggregation can influence the final configuration within a co-culture spheroid. When Janus spheroids were constructed from mouse embryonic stem (mES) cells and hepatocytes, the mES cells differentiated in a spatially distinct pattern dictated by the position of the hepatocytes. This contrasts with uniform mES differentiation observed when co-culture spheroids are formed by the conventional method of randomly mixing the two cell types. This cellular patterning method opens new possibilities for understanding and manipulating interactions between different cell types in 3D.
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