CTC Isolation from Whole Blood

PB was collected before the treatment began (baseline) and IMV was collected during surgery in order to conduct CTC evaluation. Samples were kept in plastic blood collection tubes with K2EDTA (BD Vacutainer^®, Plymouth, UK) at room temperature and were processed within 24 h of collection. The isolation procedure began with the Leucosep TM tube (bio-check LABORATORIES LTD, New Taipei City, Taiwan) to remove red blood cells from whole blood. In this step, red blood cells were removed through Ficoll–Paque PLUS (GE Healthcare Life Sciences, Taipei, Taiwan) by concentration gradient. The mononucleated cells containing CTC (peripheral blood mononuclear cells (PBMCs)) after centrifugation were collected, and the CTC cell retention rate was about 75–85% [30 (link)].