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

Analyst. 2019 Jan 28;144(3):990-996. doi: 10.1039/c8an01456f.

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.

MeSH terms

  • A549 Cells
  • Cell Adhesion*
  • Cell Separation / methods*
  • DNA, Neoplasm / analysis*
  • Genome, Human*
  • Genotype
  • HeLa Cells
  • Humans
  • Hydrogels / chemistry*
  • Single-Cell Analysis / methods*

Substances

  • DNA, Neoplasm
  • Hydrogels