title: Capture and Release of Viable Circulating Tumor Cells from Blood.
abstract: We demonstrate a method for size based capture of viable circulating tumor cell (CTC) from whole blood, along with the release of these cells from chip for downstream analysis and/or culture. The strategy employs the use of a novel Parylene C membrane slot pore microfilter to capture CTC and a coating of poly (N-iso-propylacrylamide) (PIPAAm) for thermoresponsive viable release of the captured CTC. The capture of live cells is enabled by leveraging the design of a slot pore geometry with specific dimensions to reduce the shear stress typically associated with the filtration process. While the microfilter exhibits a high capture efficiency, the release of these cells is non-trivial. Typically, only a small percentage of cells are released when techniques such as reverse flow or cell scraping are used. The strong adhesion of these epithelial cancer cells to the Parylene C membrane is attributable to non-specific electrostatic interaction. To counteract this effect, we employed the use of PIPAAm coating and exploited its thermal responsive interfacial properties to release the cells from the filter. Blood is first filtered at room temperature. Below 32 °C, PIPAAm is hydrophilic. Thereafter, the filter is placed in either culture media or a buffer maintained at 37 °C, which results in the PIPAAm turning hydrophobic, and subsequently releasing the electrostatically bound cells.
authors.affiliation: Department of Pathology, University of Miami.
authors.forename: Siddarth
authors.initials: S
authors.lastname: Rawal
authors.affiliation: SRI International.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Ao
authors.affiliation: Department of Pathology, University of Miami; Department of Biomedical Engineering, DJTMF Biomedical Nanotechnology Institute at the University of Miami, University of Miami; a.agarwal2@miami.edu.
authors.forename: Ashutosh
authors.initials: A
authors.lastname: Agarwal
mesh_terms: D002448:Cell Adhesion; D002452:Cell Count; D018929:Cell Culture Techniques; D005374:Filtration; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D009360:Neoplastic Cells, Circulating; D013314:Stress, Mechanical