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

Ryo Negishi, Reito Iwata, Tsuyoshi Tanaka, David Kisailus, Yoshiaki Maeda, Tadashi Matsunaga, Tomoko Yoshino

Research output: Contribution to journalArticle

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.

Original languageEnglish
Pages (from-to)990-996
Number of pages7
JournalAnalyst
Volume144
Issue number3
DOIs
Publication statusPublished - 2019 Feb 7

Fingerprint

Gels
gel
Hydrogel
genetic analysis
Hydrogels
Microscopes
Single-Cell Analysis
encapsulation
Encapsulation
Cell culture
Amplification
cancer
amplification
Suspensions
genome
Cell Separation
Genes
Cells
method
Genetic Heterogeneity

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy
  • Electrochemistry

Cite this

Negishi, R., Iwata, R., Tanaka, T., Kisailus, D., Maeda, Y., Matsunaga, T., & Yoshino, T. (2019). Gel-based cell manipulation method for isolation and genotyping of single-adherent cells. Analyst, 144(3), 990-996. https://doi.org/10.1039/c8an01456f

Gel-based cell manipulation method for isolation and genotyping of single-adherent cells. / Negishi, Ryo; Iwata, Reito; Tanaka, Tsuyoshi; Kisailus, David; Maeda, Yoshiaki; Matsunaga, Tadashi; Yoshino, Tomoko.

In: Analyst, Vol. 144, No. 3, 07.02.2019, p. 990-996.

Research output: Contribution to journalArticle

Negishi, R, Iwata, R, Tanaka, T, Kisailus, D, Maeda, Y, Matsunaga, T & Yoshino, T 2019, 'Gel-based cell manipulation method for isolation and genotyping of single-adherent cells' Analyst, vol. 144, no. 3, pp. 990-996. https://doi.org/10.1039/c8an01456f
Negishi, Ryo ; Iwata, Reito ; Tanaka, Tsuyoshi ; Kisailus, David ; Maeda, Yoshiaki ; Matsunaga, Tadashi ; Yoshino, Tomoko. / Gel-based cell manipulation method for isolation and genotyping of single-adherent cells. In: Analyst. 2019 ; Vol. 144, No. 3. pp. 990-996.
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