RT Journal Article SR Electronic T1 High-purity Isolation for Genotyping Rare Cancer Cells from Blood Using a Microfluidic Chip Cell Sorter JF Anticancer Research JO Anticancer Res FD International Institute of Anticancer Research SP 407 OP 417 DO 10.21873/anticanres.15499 VO 42 IS 1 A1 MIO IKEDA A1 YASUHIRO KOH A1 JUN OYANAGI A1 SHUNSUKE TERAOKA A1 MASAYUKI ISHIGE A1 YUU FUJIMURA A1 KAZUO TAKEDA A1 NAHOMI TOKUDOME A1 YUICHI OZAWA A1 HIROKI UEDA A1 NOBUYUKI YAMAMOTO YR 2022 UL http://ar.iiarjournals.org/content/42/1/407.abstract AB Background/Aim: 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. Materials and Methods: After only hemolysis and cell staining, cancer cells are enriched by repetitive sorting (3×) based on nuclear-positive, cytokeratin-positive, and CD45-negative expression. Results: 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. Conclusion: This novel isolation method applicable for preserved samples with sufficient recovery and purity may be substantially beneficial for recovering cells for subsequent molecular analysis.