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Gel-based cell manipulation method for isolation and genotyping of single-adherent cells

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Abstract

Genetic analysis of single-cells is widely recognized as a powerful tool for understanding cellular heterogeneity and obtaining genetic information from rare populations. Recently, many kinds of single-cell isolation systems have been developed to facilitate single-cell genetic analysis. However, these systems mainly target non-adherent cells or cells in a cell suspension. Thus, it is still challenging to isolate single-adherent cells of interest from a culture dish using a microscope. We had previously developed a single-cell isolation technique termed “gel-based cell manipulation” (GCM). In GCM, single-cells could be visualized by photopolymerizable-hydrogel encapsulation that made it easier to isolate the single-cells. In this study, GCM-based isolation of single-adherent cancer cells from a culture dish was demonstrated. Single-adherent cells were encapsulated in a photopolymerizable hydrogel using a microscope and isolated with high efficiency. Furthermore, whole genome amplification and sequencing for the isolated single-adherent cell could be achieved. We propose that the GCM-based approach demonstrated in this study has the potential for efficient analysis of single-adherent cells at the genetic level.

Graphical abstract: Gel-based cell manipulation method for isolation and genotyping of single-adherent cells

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Publication details

The article was received on 31 Jul 2018, accepted on 30 Sep 2018 and first published on 05 Oct 2018


Article type: Paper
DOI: 10.1039/C8AN01456F
Citation: Analyst, 2018, Advance Article
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    Gel-based cell manipulation method for isolation and genotyping of single-adherent cells

    R. Negishi, R. Iwata, T. Tanaka, D. Kisailus, Y. Maeda, T. Matsunaga and T. Yoshino, Analyst, 2018, Advance Article , DOI: 10.1039/C8AN01456F

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