title: Semiautomated isolation and molecular characterisation of single or highly purified tumour cells from CellSearch enriched blood samples using dielectrophoretic cell sorting.
abstract: BACKGROUND
  Molecular characterisation of single circulating tumour cells (CTCs) holds considerable promise for predictive biomarker assessment and to explore CTC heterogeneity. We evaluate a new method, the DEPArray system, that allows the dielectrophoretic manipulation and isolation of single and 100% purified groups of CTCs from pre-enriched blood samples and explore the feasibility of their molecular characterisation.
  
  
  METHODS
  Samples containing known numbers of two cell populations were used to assess cell loss during sample loading. Cultured breast cancer cells were isolated from spiked blood samples using CellSearch CTC and Profile kits. Single tumour cells and groups of up to 10 tumour cells were recovered with the DEPArray system and subjected to transcriptional and mutation analysis.
  
  
  RESULTS
  On average, 40% cell loss was observed when loading samples to the DEPArray system. Expected mutations in clinically relevant markers could be obtained for 60% of single recovered tumour cells and all groups of tumour cells. Reliable gene expression profiles were obtained from single cells and groups of up to 10 cells for 2 out of 3 spiked breast cancer cell lines.
  
  
  CONCLUSIONS
  We describe a semiautomated workflow for the isolation of small groups of 1 to 10 tumour cells from whole blood samples and provide proof of principle for the feasibility of their comprehensive molecular characterisation.
authors.affiliation: Translational Cancer Research Unit, Oncology Center GZA Hospitals Sint-Augustinus, Oosterveldlaan 24, Antwerp B-2610, Belgium.
authors.forename: D J E
authors.initials: DJ
authors.lastname: Peeters
authors.forename: B
authors.initials: B
authors.lastname: De Laere
authors.forename: G G
authors.initials: GG
authors.lastname: Van den Eynden
authors.forename: S J
authors.initials: SJ
authors.lastname: Van Laere
authors.forename: F
authors.initials: F
authors.lastname: Rothé
authors.forename: M
authors.initials: M
authors.lastname: Ignatiadis
authors.forename: A M
authors.initials: AM
authors.lastname: Sieuwerts
authors.forename: D
authors.initials: D
authors.lastname: Lambrechts
authors.forename: A
authors.initials: A
authors.lastname: Rutten
authors.forename: P A
authors.initials: PA
authors.lastname: van Dam
authors.forename: P
authors.initials: P
authors.lastname: Pauwels
authors.forename: M
authors.initials: M
authors.lastname: Peeters
authors.forename: P B
authors.initials: PB
authors.lastname: Vermeulen
authors.forename: L Y
authors.initials: LY
authors.lastname: Dirix
mesh_terms: D014408:Biomarkers, Tumor / Q000097:blood* / Q000235:genetics; D001943:Breast Neoplasms / Q000097:blood* / Q000235:genetics / Q000473:pathology; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D009154:Mutation / Q000235:genetics; D009360:Neoplastic Cells, Circulating / Q000473:pathology*
chemical_list: D014408:Biomarkers, Tumor