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Microsieves for the detection of circulating tumor cells in leukapheresis product in non-small cell lung cancer patients

Tamminga, M., Oomens, L., Hiltermann, T. J. N., Andree, K. C., Tibbe, A., Broekmaat, J., Schuuring, E., Terstappen, L. W. M. M. & Groen, H. J. M., Aug-2020, In : Translational lung cancer research. 9, 4, p. 1093-1100 10 p.

Research output: Contribution to journalArticleAcademicpeer-review

Background: Circulating tumor cells (CTC) in non-small cell lung cancer (NSCLC) patients are a prognostic and possible therapeutic marker, but have a low frequency of appearance. Diagnostic leukapheresis (DLA) concentrates CTC and mononuclear cells from the blood. We evaluated a protocol using two VyCAP microsieves to filter DLA product of NSCLC patients and enumerate CTC, compared with CellSearch as a gold standard. Methods: DLA was performed in NSCLC patients before starting treatment. DLA product equaling 2×108 leukocytes was diluted to 9 mL with CellSearch dilution buffer in a Transfix CTC tube. Within 72 hours the sample was filtered with a 7 μm pore microsieve and subsequently over a 5μm pore microsieve. CTC were defined as nucleated cells which stained for cytokeratin, but lacked CD45 and CD16. CellSearch detected CTC in the same volume of DLA. Results: Of 29 patients a median of 1.4 mL DLA product (range, 0.5-4.1) was filtered (2% of total product) successfully in 93% and 45% of patients using 7 and 5 μm pores, respectively. Two DLA products were unevaluable for CTC detection. Clogging of the 5 μm but not 7 μm microsieves was positively correlated with fixation time (ρ=0.51, P<0.01). VyCAP detected CTC in 44% (12/27) of DLA products. Median CTC count per mL DLA was 0 [interquartile range (IQR): 0-1]. CellSearch detected CTC in 63% of DLA products (median =0.9 CTC per mL DLA, IQR: 0-2.1). CTC counts detected by CellSearch were significantly higher compared with VyCAP (P=0.05). Conclusions: VyCAP microsieves can identify CTC in DLA product, but workflows need to be optimized.

Original languageEnglish
Pages (from-to)1093-1100
Number of pages10
JournalTranslational lung cancer research
Volume9
Issue number4
Publication statusPublished - Aug-2020

    Keywords

  • Diagnostic leukapheresis (DLA), circulating tumor cell (CTC), non-small lung cancer (NSCLC), liquid biopsy, biomarker, VyCAP microsieves, DIAGNOSTIC LEUKAPHERESIS

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