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一种用于筛选未分化人胚胎干细胞及分析的微流控方法

A Microfluidic Method for the Selection of Undifferentiated Human Embryonic Stem Cells and Analysis.

作者信息

Jabart E, Rangarajan S, Lieu C, Hack J, Conboy I, Sohn L L

机构信息

Dept. of Bioengineering, University of California, Berkeley 94720, USA.

School of Medicine, Creighton University, Omaha, NE 68178, USA.

出版信息

Microfluid Nanofluidics. 2015 May;18(5-6):955-966. doi: 10.1007/s10404-014-1485-9. Epub 2014 Sep 28.

Abstract

Conventional cell-sorting methods such as fluorescence-activated cell sorting (FACS) or magnetic-activated cell sorting (MACS) can suffer from certain shortcomings such as lengthy sample preparation time, cell modification through antibody labeling, and cell damage due to exposure to high shear forces or to attachment of superparamagnetic Microbeads. In light of these drawbacks, we have recently developed a label-free, microfluidic platform that can not only select cells with minimal sample preparation but also enable analysis of cells in situ. We demonstrate the utility of our platform by successfully isolating undifferentiated human embryonic stem cells (hESCs) from a heterogeneous population based on the undifferentiated stem-cell marker SSEA-4. Importantly, we show that, in contrast to MACS or FACS, cells isolated by our method have very high viability (~90%). Overall, our platform technology could likely be applied to other cell types beyond hESCs and to a variety of heterogeneous cell populations in order to select and analyze cells of interest.

摘要

传统的细胞分选方法,如荧光激活细胞分选(FACS)或磁激活细胞分选(MACS),可能存在某些缺点,例如样品制备时间长、通过抗体标记对细胞进行修饰以及由于暴露于高剪切力或超顺磁性微珠附着而导致的细胞损伤。鉴于这些缺点,我们最近开发了一种无标记的微流控平台,该平台不仅可以在最少样品制备的情况下选择细胞,还能够对原位细胞进行分析。我们通过基于未分化干细胞标志物SSEA-4从异质群体中成功分离出未分化的人类胚胎干细胞(hESCs),证明了我们平台的实用性。重要的是,我们表明,与MACS或FACS不同,通过我们的方法分离的细胞具有非常高的活力(约90%)。总体而言,我们的平台技术可能适用于hESCs以外的其他细胞类型以及各种异质细胞群体,以便选择和分析感兴趣的细胞。

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