Diffusion-based extraction of DMSO from a cell suspension in a three stream, vertical microchannel.
Diffusion-based extraction of DMSO from a cell suspension in a three stream, vertical microchannel.
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DOI：
10.1002/bit.24499
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发表时间：
2012-09
影响因子：
3.8
通讯作者：
Lemke E
中科院分区：
文献类型：
--
作者：
Hanna J;Hubel A;Lemke E
细胞通常被冷冻保存以用于研究和治疗应用。最常见的冷冻保护剂 (CPA) 二甲基亚砜 (DMSO) 有毒，在使用细胞之前必须将其去除。这项研究使用了一种微流体装置，其中三股流垂直平行地流过深度为 500 μm 的矩形通道。两条洗涤液流在充满 DMSO 的细胞流的两侧流动，使 DMSO 扩散到洗涤液中并被去除，并收集洗涤后的样品。研究了设备从细胞流中提取 DMSO 的能力，样品流速为 0.5 至 4.0 mL/min (Pe = 1,263–10,100)。使用悬浮液中的 Jurkat 细胞（淋巴细胞）对从设备中回收细胞进行了研究，细胞体积百分比范围为 0.5% 至 15%。在所有研究条件下，细胞回收率均 > 95%，而 DMSO 去除率可与之前开发的双流装置相媲美，只需四分之一的装置长度或四倍的流速即可实现。与标准细胞清洗技术相比，高细胞回收率提高了约 25%，并且实现的高流速在微流体装置中并不常见，从而可以处理临床相关的细胞群。
Cells are routinely cryopreserved for investigative and therapeutic applications. The most common cryo-protective agent (CPA), dimethyl sulfoxide (DMSO), is toxic, and must be removed before cells can be used. This study uses a microfluidic device in which three streams flow vertically in parallel through a rectangular channel 500 μm in depth. Two wash streams flow on either side of a DMSO-laden cell stream, allowing DMSO to diffuse into the wash and be removed, and the washed sample to be collected. The ability of the device to extract DMSO from a cell stream was investigated for sample flow rates from 0.5 to 4.0 mL/min (Pe = 1,263–10,100). Recovery of cells from the device was investigated using Jurkat cells (lymphoblasts) in suspensions ranging from 0.5% to 15% cells by volume. Cell recovery was >95% for all conditions investigated, while DMSO removal comparable to a previously developed two-stream device was achieved in either one-quarter the device length, or at four times the flow rate. The high cell recovery is a ~25% improvement over standard cell washing techniques, and high flow rates achieved are uncommon among microfluidic devices, allowing for processing of clinically relevant cell populations.
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