1. Gel-based cell manipulation method for isolation and genotyping of single-adherent cells
- Author
-
Tsuyoshi Tanaka, Reito Iwata, Yoshiaki Maeda, Tadashi Matsunaga, Ryo Negishi, Tomoko Yoshino, and David Kisailus
- Subjects
Genotype ,Cell ,Cell Separation ,02 engineering and technology ,Computational biology ,Biology ,01 natural sciences ,Biochemistry ,Genome ,Analytical Chemistry ,law.invention ,law ,Cell Adhesion ,Electrochemistry ,medicine ,Humans ,Environmental Chemistry ,Cell adhesion ,Genotyping ,Spectroscopy ,A549 cell ,Whole Genome Amplification ,Genome, Human ,Petri dish ,010401 analytical chemistry ,Hydrogels ,DNA, Neoplasm ,021001 nanoscience & nanotechnology ,0104 chemical sciences ,medicine.anatomical_structure ,A549 Cells ,Cancer cell ,Single-Cell Analysis ,0210 nano-technology ,HeLa 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.
- Published
- 2019
- Full Text
- View/download PDF