CellMag-CARWash: A High Throughput Droplet Microfluidic Device for Live Cell Isolation and Single Cell Applications

IF 3.2 3区 生物学 Q3 MATERIALS SCIENCE, BIOMATERIALS
Brittany T. Rupp, Claire D. Cook, Emma A. Purcell, Matei Pop, Abigail E. Radomski, Nicolas Mesyngier, Ryan C. Bailey, Sunitha Nagrath
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Abstract

The recent push toward understanding an individual cell's behavior and identifying cellular heterogeneity has created an unmet need for technologies that can probe live cells at the single-cell level. Cells within a population are known to exhibit heterogeneous responses to environmental cues. These differences can lead to varied cellular states, behavior, and responses to therapeutics. Techniques are needed that are not only capable of processing and analyzing cellular populations at the single cell level, but also have the ability to isolate specific cell populations from a complex sample at high throughputs. The new CellMag-Coalesce-Attract-Resegment Wash (CellMag-CARWash) system combines positive magnetic selection with droplet microfluidic devices to isolate cells of interest from a mixture with >93% purity and incorporate treatments within individual droplets to observe single cell biological responses. This workflow is shown to be capable of probing the single cell extracellular vesicle (EV) secretion of MCF7 GFP cells. This article reports the first measurement of β-Estradiol's effect on EV secretion from MCF7 cells at the single cell level. Single cell processing revealed that MCF7 GFP cells possess a heterogeneous response to β-Estradiol stimulation with a 1.8-fold increase relative to the control.

Abstract Image

CellMag-CARWash:用于活细胞分离和单细胞应用的高通量液滴微流控装置
最近,人们正努力了解单个细胞的行为并识别细胞的异质性,这就产生了对能在单细胞水平上探测活细胞的技术的需求。众所周知,群体中的细胞对环境线索的反应各不相同。这些差异会导致不同的细胞状态、行为和对疗法的反应。我们需要的技术不仅能在单细胞水平上处理和分析细胞群,还能以高通量从复杂样本中分离出特定细胞群。新型 CellMag-Coalesce-Attract-Resegment Wash(CellMag-CARWash)系统将正磁性选择与液滴微流控装置相结合,从纯度大于 93% 的混合物中分离出感兴趣的细胞,并在单个液滴中进行处理,观察单细胞的生物反应。该工作流程能够探测 MCF7 GFP 细胞的单细胞胞外囊泡 (EV) 分泌。本文首次在单细胞水平上测量了β-雌二醇对MCF7细胞EV分泌的影响。单细胞处理显示,MCF7 GFP细胞对β-雌二醇刺激具有异质性反应,相对于对照组增加了1.8倍。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Advanced biology
Advanced biology Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)
CiteScore
6.60
自引率
0.00%
发文量
130
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