Purification of cell subpopulations via immiscible filtration assisted by surface tension (IFAST).
Purification of cell subpopulations via immiscible filtration assisted by surface tension (IFAST).
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DOI：
10.1007/s10544-011-9573-z
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发表时间：
2011-12
影响因子：
2.8
通讯作者：
Beebe, David J.
中科院分区：
文献类型：
--
作者：
Berry, Scott M.;Strotman, Lindsay N.;Kueck, Jessica D.;Alarid, Elaine T.;Beebe, David J.
从更大的混合细胞群中选择性分离细胞亚群是许多生物医学测定的关键准备过程。在这里，我们提出了一种新的细胞分离平台，与现有设备相比具有一系列独特的优势。我们的技术被称为表面张力辅助的不混溶过滤，利用与微尺度相关的物理现象来建立由不混溶液体组成的流体屏障。通过免疫捕获将磁响应颗粒附着到目标细胞群上，我们可以选择性地将这个细胞群转运穿过不混溶的屏障并进入单独的水溶液中。与不混溶相/水相边界相关的高界面能可防止不需要的细胞或其他污染物无意中穿过不混溶相。我们已经证明，使用荧光颗粒、基质细胞和全血作为“背景”，我们可以成功分离出约 70% 的目标乳腺癌细胞群，平均纯度 >80%。通过串联连接两个不混溶的屏障可以获得更高的纯度，这一修改仅略微增加了操作复杂性。此外，可以以近乎瞬时的方式平行批次处理多个样品，无需任何清洗，清洗可能会导致靶细胞稀释（阴性选择）或显着不受控制的损失（阳性选择）。最后，观察到细胞在经历不混溶阶段后仍保持活力和增殖，表明该过程适用于各种下游测定，包括那些需要完整活细胞的测定。
The selective isolation of a sub-population of cells from a larger, mixed population is a critical preparatory process to many biomedical assays. Here, we present a new cell isolation platform with a unique set of advantages over existing devices. Our technology, termed Immiscible Filtration Assisted by Surface Tension, exploits physical phenomena associated with the microscale to establish fluidic barriers composed of immiscible liquids. By attaching magnetically-responsive particles to a target cell population via immunocapture, we can selectively transport this population across the immiscible barrier and into a separate aqueous solution. The high interfacial energy associated with the immiscible phase / aqueous phase boundaries prevents unwanted cells or other contaminants from inadvertently crossing the immiscible phase. We have demonstrated, using fluorescent particles, stromal cells, and whole blood as “background”, that we can successfully isolate ~70% of a target breast cancer cell population with an average purity of >80%. Increased purity was obtained by coupling two immiscible barriers in series, a modification that only slightly increases operational complexity. Furthermore, several samples can be processed in parallel batches in a near-instantaneous manner without the requirement of any washing, which can cause dilution (negative selection) or significant uncontrolled loss (positive selection) of target cells. Finally, cells were observed to remain viable and proliferative following traverse through the immiscible phase, indicating that this process is suitable for a variety of downstream assays, including those requiring intact living cells.
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影响因子：
2.4
作者：
Tsutsui H;Ho CM
通讯作者：
Ho CM
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作者：
MacDonald, MP;Spalding, GC;Dholakia, K
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Dholakia, K
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Zhao, B;Moore, JS;Beebe, DJ
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Beebe, DJ
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Chen, Hong;Abolmatty, Assem;Faghri, Mohammad
通讯作者：
Faghri, Mohammad
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作者：
Huang, LR;Cox, EC;Sturm, JC
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Sturm, JC