Manipulation of a Single Circulating Tumor Cell Using Visualization of Hydrogel Encapsulation toward Single-Cell Whole-Genome Amplification.
Genetic characterization of circulating tumor cells (CTCs) could guide the choice of therapies for individual patients and also facilitate the development of new drugs. We previously developed a CTC recovery system using a microcavity array, which demonstrated highly efficient CTC recovery based on differences in cell size and deformability. However, the CTC recovery system lacked an efficient cell manipulation tool suitable for subsequent genetic analysis. Here, we resolve this issue and present a simple and rapid manipulation method for single CTCs using a photopolymerized hydrogel, polyethylene glycol diacrylate (PEGDA), which is useful for subsequent genetic analysis. First, PEGDA was introduced into the cells entrapped on the microcavity array. Then, excitation light was projected onto the target single cells for encapsulation of each CTC by confocal laser-scanning microscopy. The encapsulated single CTCs could be visualized by the naked eye and easily handled with tweezers. The single CTCs were only partially encapsulated on the PEGDA hydrogel, which allowed for sufficient whole-genome amplification and accurate genotyping. Our proposed methodology is a valuable tool for the rapid and simple manipulation of single CTCs and is expected to become widely utilized for analyses of mammalian cells and microorganisms in addition to CTCs.
Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology , 2-24-16, Naka-cho, Koganei, Tokyo, 184-8588, Japan.
Tomoko
T
Yoshino
Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology , 2-24-16, Naka-cho, Koganei, Tokyo, 184-8588, Japan.
Tsuyoshi
T
Tanaka
Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology , 2-24-16, Naka-cho, Koganei, Tokyo, 184-8588, Japan.
Seita
S
Nakamura
Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology , 2-24-16, Naka-cho, Koganei, Tokyo, 184-8588, Japan.
Ryo
R
Negishi
Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology , 2-24-16, Naka-cho, Koganei, Tokyo, 184-8588, Japan.
Masahito
M
Hosokawa
Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology , 2-24-16, Naka-cho, Koganei, Tokyo, 184-8588, Japan.
Tadashi
T
Matsunaga
D045744:Cell Line, Tumor; D004273:DNA, Neoplasm / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D066246:ErbB Receptors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015894:Genome, Human; D005838:Genotype; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D008027:Light; D018613:Microscopy, Confocal; D009360:Neoplastic Cells, Circulating / Q000737:chemistry* / Q000378:metabolism / Q000473:pathology; D058105:Polymerization / Q000528:radiation effects; D017422:Sequence Analysis, DNA; D059010:Single-Cell Analysis
D004273:DNA, Neoplasm; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; C512478:EGFR protein, human; D066246:ErbB Receptors