Gel-based cell manipulation method for isolation and genotyping of single-adherent cells. Issue 3 (10th October 2018)

• Record Type: Journal Article
• Authors: Negishi, Ryo; Iwata, Reito; Tanaka, Tsuyoshi; Kisailus, David; Maeda, Yoshiaki; Matsunaga, Tadashi; Yoshino, Tomoko
• Is Part Of: Analyst. Volume 144:Issue 3(2019)
• Page Start: 990
• Page End: 996
• Publication Date: 2018-10-10
• Subjects: Chemistry, Analytic -- Periodicals; 543
• DOI: 10.1039/c8an01456f
• Languages: English
• ISSNs: 0003-2654

Abstract

Abstract : The simple and rapid method for isolation of single-adherent cells from a culture dish was developed and applied to genetic analysis of single-cells. 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.