Microfluidic Sorting of Cells by Viability Based on Differences in Cell Stiffness.
Microfluidic Sorting of Cells by Viability Based on Differences in Cell Stiffness.
复制标题
DOI：
10.1038/s41598-017-01807-z
复制
发表时间：
2017-05-17
影响因子：
4.6
通讯作者：
Sulchek T
中科院分区：
文献类型：
--
作者：
Islam M;Brink H;Blanche S;DiPrete C;Bongiorno T;Stone N;Liu A;Philip A;Wang G;Lam W;Alexeev A;Waller EK;Sulchek T
活细胞的富集是获得用于细胞治疗的有效产品的必要步骤。虽然存在表征细胞活力的程序，但排除非活细胞的大多数方法需要使用密度梯度离心或使用细胞活力的分子标记的基于抗体的细胞分选。我们报告了一种无标记的微流控技术，利用细胞硬度的差异来分离活细胞和死细胞。该装置使用具有相对于细胞流动方向呈对角线的重复脊的通道。被引导通过通道的刚性非活细胞被压缩并垂直于通道长度平移，而软活细胞遵循流体动力学流动。作为概念的证明，Jurkat细胞被富集到高纯度的活细胞的185倍。细胞硬度被验证为分选参数，因为非活细胞比活细胞硬得多。为了突出造血干细胞移植的实用性，将冷冻的脐带血样品解冻，并且活的有核细胞的纯度从65%增加到94%以上，活细胞的回收率为73%。因此，微流体刚度分选可以简单有效地获得高纯度的活细胞群。
The enrichment of viable cells is an essential step to obtain effective products for cell therapy. While procedures exist to characterize the viability of cells, most methods to exclude nonviable cells require the use of density gradient centrifugation or antibody-based cell sorting with molecular labels of cell viability. We report a label-free microfluidic technique to separate live and dead cells that exploits differences in cellular stiffness. The device uses a channel with repeated ridges that are diagonal with respect to the direction of cell flow. Stiff nonviable cells directed through the channel are compressed and translated orthogonally to the channel length, while soft live cells follow hydrodynamic flow. As a proof of concept, Jurkat cells are enriched to high purity of viable cells by a factor of 185-fold. Cell stiffness was validated as a sorting parameter as nonviable cells were substantially stiffer than live cells. To highlight the utility for hematopoietic stem cell transplantation, frozen samples of cord blood were thawed and the purity of viable nucleated cells was increased from 65% to over 94% with a recovery of 73% of the viable cells. Thus, the microfluidic stiffness sorting can simply and efficiently obtain highly pure populations of viable cells.