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An automated microfluidic device for assessment of mammalian cell genetic stability

renchunxiao 添加于 2012/7/24 14:41:17  1124次阅读 | 0次推荐 | 0个评论

Single-cell transcriptome contains reliable gene regulatory relationships because gene–gene interactions only happen within a mammalian cell. While the study of gene–gene interactions enables us to understand the molecular mechanism of cellular events and evaluate molecular characteristics of a mammalian cell population, its complexity requires an analysis of a large number of single-cells at various stages. However, many existing microfluidic platforms cannot process single-cells effectively for routine molecular analysis. To address these challenges, we develop an integrated system with individual controller for effective single-cell transcriptome analysis. In this paper, we report an integrated microfluidic approach to rapidly measure gene expression in individual cells for genetic stability assessment of a cell population. Inside this integrated microfluidic device, the cells are individually manipulated and isolated in an array using micro sieve structures, then transferred into different nanoliter reaction chambers for parallel processing of single-cell transcriptome analysis. This device enables us to manipulate individual single-cells into nanoliter reactor with high recovery rate. We have performed gene expression analysis for a large number of HeLa cells and 293T cells expanded from a single-cell. Our data shows that even the house-keeping genes are expressed at heterogeneous levels within a clone of cells. The heterogeneity of actin expression reflects the genetic stability, and the expression distribution is different between cancer cells (HeLa) and immortalized 293T cells. The result demonstrates that this platform has the potential for assessment of genetic stability in cancer diagnosis.

作 者:Yan Chen , Baoyue Zhang , Hongtao Feng , Weiliang Shu , Gina Y. Chen and Jiang F. Zhong
期刊名称: lab on a chip
期卷页: 第卷 第期 页
学科领域:信息科学 » 计算机科学 » 计算机科学的基础理论
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原文链接:http://pubs.rsc.org/en/content/articlelanding/2012/lc/c2lc40437k
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相关报道: http://paper.sciencenet.cn/htmlpaper/201272414424166125192.shtm
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