Transwells with Microstamped Membranes Produce Micropatterned Two-Dimensional and Three-Dimensional Co-Cultures
Transwells with Microstamped Membranes Produce Micropatterned Two-Dimensional and Three-Dimensional Co-Cultures
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DOI：
10.1089/ten.tec.2010.0305
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发表时间：
2011-01-01
影响因子：
3
通讯作者：
Takayama, Shuichi
中科院分区：
文献类型：
--
作者：
Torisawa, Yu-Suke;Mosadegh, Bobak;Takayama, Shuichi
这篇文章描述了一种简单而快速的细胞图案化方法，以在商业上可用的Trswell中形成共培养微阵列。使用聚二甲基硅氧烷(PDMS)印章在Transwell的底面上印刷一层薄的聚二甲基硅氧烷(PDMS)。根据薄层PDMS层的几何特征，通过流体动力将细胞引导到膜上，仅在未涂覆PDMS的区域上生成任意细胞图案。根据滤膜的表面处理，可以产生表面附着细胞的微图案(我们将其称为二维)或非表面附着细胞簇(我们将其称为三维)。此外，可以通过在Transwell膜上或在Transwell的底室中引入不同类型的细胞来建立共培养。我们证明，这种共培养方法可以基于异种细胞-细胞相互作用来评估小鼠胚胎干细胞(MES)的分化。MES细胞与HepG2细胞共培养降低了MES细胞SOX17的表达，细胞与细胞直接接触进一步降低了SOX17的表达，表明与HepG2细胞共培养通过可溶性因子和细胞-细胞接触抑制内胚层分化。这种方法简单，用户友好，应该是广泛有用的研究细胞的形状和细胞之间的相互作用。
This article describes a simple and rapid cell patterning method to form co-culture microarrays in commercially available Transwells. A thin poly(dimethylsiloxane) (PDMS) layer is printed on the underside of a Transwell using a PDMS stamp. Arbitrary cellular patterns are generated according to the geometric features of the thin PDMS layer through hydrodynamic forces that guide cells onto the membrane only over the PDMS-uncoated regions. Micropatterns of surface-adhered cells (we refer to this as two-dimensional) or non-surface-adhered clusters of cells (we refer to this as three-dimensional) can be generated depending on the surface treatment of the filter membrane. Additionally, co-cultures can be established by introducing different types of cells on the membrane or in the bottom chamber of the Transwell. We show that this co-culture method can evaluate mouse embryonic stem (mES) cell differentiation based on heterogeneous cell-cell interactions. Co-culture of mES cells and HepG2 cells decreased SOX17 expression of mES cells, and direct cell-cell contact further decreased SOX17 expression, indicating that co-culture with HepG2 cells inhibits endoderm differentiation through soluble factors and cell-cell contact. This method is simple and user-friendly and should be broadly useful to study cell shapes and cell-cell interactions.