BACKGROUND: Circulating tumor cells (CTCs) can be enriched from peripheral blood by leukocyte depletion. In this study, a PowerMag system was designed to enhance leukocyte depletion and CTC detection. METHODS: The efficiency of PowerMag in leukocyte depletion and recovery of CTCs was evaluated. The cell populations in leukocyte-depleted cell filtrate were characterized by immunofluorescence staining using anti-EpCAM and anti-CD45 antibodies. The suitability of PowerMag in monitoring cancer patient CTCs was assessed by analyzing blood samples collected at baseline and 2 weeks after baseline from the patients with colorectal cancer (CRC, n=24) and head and neck squamous cell carcinoma (HNSCC, n=28). RESULTS: Only few contaminated CD45(+)-leukocytes were retained in the leukocyte-depleted cell filtrate. The enriched CTCs were viable with a recovery rate of 46-62%. Immunofluorescence staining of the nucleated cells enriched by PowerMag revealed 2 major cell populations EpCAM(+)CD45(-) and EpCAM(-)CD45(-). The number for both cell types was significantly increased in cancer patients when compared to healthy control. After chemotherapy, the number of EpCAM(+)CD45(-) and EpCAM(-)CD45(-) cells was decreased more prominently for CRC and HNSCC patients, respectively. CONCLUSIONS: PowerMag holds great promise as a platform for leukocyte depletion and CTC detection in a clinical setting.
BACKGROUND: Circulating tumor cells (CTCs) can be enriched from peripheral blood by leukocyte depletion. In this study, a PowerMag system was designed to enhance leukocyte depletion and CTC detection. METHODS: The efficiency of PowerMag in leukocyte depletion and recovery of CTCs was evaluated. The cell populations in leukocyte-depleted cell filtrate were characterized by immunofluorescence staining using anti-EpCAM and anti-CD45 antibodies. The suitability of PowerMag in monitoring cancerpatient CTCs was assessed by analyzing blood samples collected at baseline and 2 weeks after baseline from the patients with colorectal cancer (CRC, n=24) and head and neck squamous cell carcinoma (HNSCC, n=28). RESULTS: Only few contaminated CD45(+)-leukocytes were retained in the leukocyte-depleted cell filtrate. The enriched CTCs were viable with a recovery rate of 46-62%. Immunofluorescence staining of the nucleated cells enriched by PowerMag revealed 2 major cell populations EpCAM(+)CD45(-) and EpCAM(-)CD45(-). The number for both cell types was significantly increased in cancerpatients when compared to healthy control. After chemotherapy, the number of EpCAM(+)CD45(-) and EpCAM(-)CD45(-) cells was decreased more prominently for CRC and HNSCC patients, respectively. CONCLUSIONS: PowerMag holds great promise as a platform for leukocyte depletion and CTC detection in a clinical setting.
Authors: Giuseppe Galletti; Matthew S Sung; Linda T Vahdat; Manish A Shah; Steven M Santana; Giuseppe Altavilla; Brian J Kirby; Paraskevi Giannakakou Journal: Lab Chip Date: 2013-11-07 Impact factor: 6.799