Polymeric Aqueous Biphasic System Rehydration Facilitates High Throughput Cell Exclusion Patterning For Cell Migration Studies.
Polymeric Aqueous Biphasic System Rehydration Facilitates High Throughput Cell Exclusion Patterning For Cell Migration Studies.
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DOI：
10.1002/adfm.201002559
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发表时间：
2011-08-09
影响因子：
19
通讯作者：
Takayama, Shuichi
中科院分区：
文献类型：
--
作者：
Tavana, Hossein;Kaylan, Kerim;Bersano-Begey, Tommaso;Luker, Kathryn E.;Luker, Gary D.;Takayama, Shuichi
本文描述了一种由聚合物水两相体系促进的细胞排斥图像化方法。含有细胞的浸渍水相（聚乙二醇）使底物上的较密相（葡聚糖）的干燥圆盘再水合以形成葡聚糖液滴。在形成相聚合物的适当性质下，再水合液滴与浸入相保持不混相。两相体系的适当配方确保再水合液滴与周围水相之间的界面张力阻止细胞穿过界面，从而使细胞仅粘附在葡聚糖相液滴周围的底物区域上。冲洗出图案两相试剂显示细胞单层包含一个明确的圆形间隙，作为单层细胞的迁移生态位。随着时间的推移，细胞向细胞排除区域的迁移很容易被可视化和量化。这种“间隙愈合”迁移试验的96孔板格式证明了检测已知细胞骨架靶向剂对细胞迁移抑制的能力。这种简单的方法，只需要一个传统的液体处理器和容易制备的聚合物溶液，为高通量细胞迁移分析开辟了新的机会。
This paper describes a cell-exclusion patterning method facilitated by a polymeric aqueous two-phase system. The immersion aqueous phase (polyethylene glycol) containing cells rehydrates a dried disk of the denser phase (dextran) on the substrate to form a dextran droplet. With the right properties of the phase-forming polymers, the rehydrating droplet remains immiscible with the immersion phase. Proper formulation of the two-phase system ensures that the interfacial tension between the rehydrating droplet and the surrounding aqueous phase prevents cells from crossing the interface so that cells only adhere to the regions of the substrate around the dextran phase droplet. Washing out the patterning two-phase reagents reveals a cell monolayer containing a well-defined circular gap that serves as the migration niche for cells of the monolayer. Migration of cells into the cell-excluded area is readily visualized and quantified over time. A 96-well plate format of this “gap healing” migration assay demonstrates the ability to detect inhibition of cell migration by known cytoskeleton targeting agents. This straightforward method, which only requires a conventional liquid handler and readily prepared polymer solutions, opens new opportunities for high throughput cell migration assays.
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DOI：
10.1186/bcr2416
发表时间：
2009
期刊：
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影响因子：
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作者：
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通讯作者：
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DOI：
10.1039/c0ib00038h
发表时间：
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期刊：
Integrative biology : quantitative biosciences from nano to macro
影响因子：
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作者：
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影响因子：
15
作者：
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通讯作者：
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影响因子：
41.2
作者：
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影响因子：
64.8
作者：
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通讯作者：
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