title: Separable bilayer microfiltration device for viable label-free enrichment of circulating tumour cells.
abstract: The analysis of circulating tumour cells (CTCs) in cancer patients could provide important information for therapeutic management. Enrichment of viable CTCs could permit performance of functional analyses on CTCs to broaden understanding of metastatic disease. However, this has not been widely accomplished. Addressing this challenge, we present a separable bilayer (SB) microfilter for viable size-based CTC capture. Unlike other single-layer CTC microfilters, the precise gap between the two layers and the architecture of pore alignment result in drastic reduction in mechanical stress on CTCs, capturing them viably. Using multiple cancer cell lines spiked in healthy donor blood, the SB microfilter demonstrated high capture efficiency (78-83%), high retention of cell viability (71-74%), high tumour cell enrichment against leukocytes (1.7-2 × 10(3)), and widespread ability to establish cultures post-capture (100% of cell lines tested). In a metastatic mouse model, SB microfilters successfully enriched viable mouse CTCs from 0.4-0.6 mL whole mouse blood samples and established in vitro cultures for further genetic and functional analysis. Our preliminary studies reflect the efficacy of the SB microfilter device to efficiently and reliably enrich viable CTCs in animal model studies, constituting an exciting technology for new insights in cancer research.
authors.affiliation: Micro &Nano Integrated Biosystem (MINIBio) Laboratory, Department of Biomedical Engineering and Materials Research Institute, Pennsylvania State University, University Park, PA 16802, U.S.A.
authors.forename: Ming-Da
authors.initials: MD
authors.lastname: Zhou
authors.affiliation: Micro &Nano Integrated Biosystem (MINIBio) Laboratory, Department of Biomedical Engineering and Materials Research Institute, Pennsylvania State University, University Park, PA 16802, U.S.A.
authors.forename: Sijie
authors.initials: S
authors.lastname: Hao
authors.affiliation: 1] Department of Pathology, University of Miami - Miller School of Medicine, Miami, FL 33136, U.S.A. [2] Sheila and David Fuente Graduate Program in Cancer Biology, University of Miami - Miller School of Medicine, Miami, FL 33136, U.S.A. [3] Dr John T Macdonald Foundation Biomedical Nanotechnology Institute, University of Miami - Miller School of Medicine, Miami, FL 33136, U.S.A.
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Williams
authors.affiliation: Micro &Nano Integrated Biosystem (MINIBio) Laboratory, Department of Biomedical Engineering and Materials Research Institute, Pennsylvania State University, University Park, PA 16802, U.S.A.
authors.forename: Ramdane A
authors.initials: RA
authors.lastname: Harouaka
authors.affiliation: 1] Sheila and David Fuente Graduate Program in Cancer Biology, University of Miami - Miller School of Medicine, Miami, FL 33136, U.S.A. [2] Department of Microbiology and Immunology, University of Miami - Miller School of Medicine, Miami, FL 33136, U.S.A.
authors.forename: Brett
authors.initials: B
authors.lastname: Schrand
authors.affiliation: 1] Department of Pathology, University of Miami - Miller School of Medicine, Miami, FL 33136, U.S.A. [2] Dr John T Macdonald Foundation Biomedical Nanotechnology Institute, University of Miami - Miller School of Medicine, Miami, FL 33136, U.S.A.
authors.forename: Siddarth
authors.initials: S
authors.lastname: Rawal
authors.affiliation: 1] Department of Pathology, University of Miami - Miller School of Medicine, Miami, FL 33136, U.S.A. [2] Sheila and David Fuente Graduate Program in Cancer Biology, University of Miami - Miller School of Medicine, Miami, FL 33136, U.S.A. [3] Dr John T Macdonald Foundation Biomedical Nanotechnology Institute, University of Miami - Miller School of Medicine, Miami, FL 33136, U.S.A.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Ao
authors.affiliation: 1] Sheila and David Fuente Graduate Program in Cancer Biology, University of Miami - Miller School of Medicine, Miami, FL 33136, U.S.A. [2] Department of Microbiology and Immunology, University of Miami - Miller School of Medicine, Miami, FL 33136, U.S.A.
authors.forename: Randall
authors.initials: R
authors.lastname: Brenneman
authors.affiliation: Department of Microbiology and Immunology, University of Miami - Miller School of Medicine, Miami, FL 33136, U.S.A.
authors.forename: Eli
authors.initials: E
authors.lastname: Gilboa
authors.affiliation: Caltech Micromachining Laboratory, California Institute of Technology, MC 136-93, Pasadena, CA 91125, U.S.A.
authors.forename: Bo
authors.initials: B
authors.lastname: Lu
authors.affiliation: Department of Pharmaceutical Sciences, Washington State University College of Pharmacy, Spokane, WA 99210, U.S.A.
authors.forename: Shuwen
authors.initials: S
authors.lastname: Wang
authors.affiliation: Department of Pharmaceutical Sciences, Washington State University College of Pharmacy, Spokane, WA 99210, U.S.A.
authors.forename: Jiyue
authors.initials: J
authors.lastname: Zhu
authors.affiliation: 1] Department of Pathology, University of Miami - Miller School of Medicine, Miami, FL 33136, U.S.A. [2] Dr John T Macdonald Foundation Biomedical Nanotechnology Institute, University of Miami - Miller School of Medicine, Miami, FL 33136, U.S.A.
authors.forename: Ram
authors.initials: R
authors.lastname: Datar
authors.affiliation: 1] Department of Pathology, University of Miami - Miller School of Medicine, Miami, FL 33136, U.S.A. [2] Dr John T Macdonald Foundation Biomedical Nanotechnology Institute, University of Miami - Miller School of Medicine, Miami, FL 33136, U.S.A.
authors.forename: Richard
authors.initials: R
authors.lastname: Cote
authors.affiliation: Caltech Micromachining Laboratory, California Institute of Technology, MC 136-93, Pasadena, CA 91125, U.S.A.
authors.forename: Yu-Chong
authors.initials: YC
authors.lastname: Tai
authors.affiliation: Micro &Nano Integrated Biosystem (MINIBio) Laboratory, Department of Biomedical Engineering and Materials Research Institute, Pennsylvania State University, University Park, PA 16802, U.S.A.
authors.forename: Si-Yang
authors.initials: SY
authors.lastname: Zheng
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000295:instrumentation* / Q000379:methods*; D002470:Cell Survival; D004195:Disease Models, Animal; D006801:Humans; D051379:Mice; D008851:Micropore Filters; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D015203:Reproducibility of Results