Department of Chemical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA.
Biointerfaces Institute, University of Michigan, Ann Arbor, MI, 48109, USA.
Adv Biol (Weinh). 2024 Jul;8(7):e2400066. doi: 10.1002/adbi.202400066. Epub 2024 May 13.
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.
近年来,人们越来越关注单个细胞的行为,并识别细胞异质性,这就产生了对能够在单细胞水平探测活细胞的技术的未满足需求。已知细胞群体中的细胞对环境信号表现出异质响应。这些差异会导致不同的细胞状态、行为以及对治疗的反应。需要开发不仅能够在单细胞水平处理和分析细胞群体,而且还能够以高通量从复杂样本中分离特定细胞群体的技术。新的 CellMag-Coalesce-Attract-Resegment Wash(CellMag-CARWash)系统将正磁选与液滴微流控装置相结合,从混合物中以>93%的纯度分离出感兴趣的细胞,并在单个液滴中加入处理,以观察单个细胞的生物学反应。该工作流程可用于探测 MCF7 GFP 细胞的单个细胞细胞外囊泡(EV)分泌。本文首次报道了在单细胞水平测量β-雌二醇对 MCF7 细胞 EV 分泌的影响。单细胞处理显示,与对照相比,MCF7 GFP 细胞对β-雌二醇刺激的反应具有异质性,增加了 1.8 倍。