High-purity Isolation for Genotyping Rare Cancer Cells from Blood Using a Microfluidic Chip Cell Sorter
| Autor: | Mio, Ikeda, Yasuhiro, Koh, Jun, Oyanagi, Shunsuke, Teraoka, Masayuki, Ishige, Yuu, Fujimura, Kazuo, Takeda, Nahomi, Tokudome, Yuichi, Ozawa, Hiroki, Ueda, Nobuyuki, Yamamoto |
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| Rok vydání: | 2021 |
| Předmět: | |
| Zdroj: | Anticancer Research. 42:407-417 |
| ISSN: | 1791-7530 0250-7005 |
| Popis: | A multistep sorting method for enrichment of rare cells, such as circulating tumor cells, in the blood without cumbersome pretreatments required by most flow cytometry-based methods, which lead to high cost and decreased detection efficiency, was developed.After only hemolysis and cell staining, cancer cells are enriched by repetitive sorting (3×) based on nuclear-positive, cytokeratin-positive, and CD45-negative expression.Experiments using spikes of PC-9 cells showed a mean recovery of 65% and mean purity of 83%, which was retained up to 72 hours after blood draw using preservative tubes. Significant differences in expression level of programmed death-ligand 1 or vimentin were observed between high- and low-expressing cell lines, concurrently with enrichment. Next-generation sequencing analysis of recovered PC-9, A549, and MDA-MB231 cells successfully detected all known mutations.This novel isolation method applicable for preserved samples with sufficient recovery and purity may be substantially beneficial for recovering cells for subsequent molecular analysis. |
| Databáze: | OpenAIRE |
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