title: 3D porous chitosan-alginate scaffolds: a new matrix for studying prostate cancer cell-lymphocyte interactions in vitro.
abstract: The treatment of castration-resistant prostate cancer (CRPC) remains palliative. Immunotherapy offers a potentially effective therapy for CRPC; however, its advancement into the clinic has been slow, in part because of the lack of representative in vitro tumor models that resemble the in vivo tumor microenvironment for studying interactions of CRPC cells with immune cells and other potential therapeutics. This study evaluates the use of 3D porous chitosan-alginate (CA) scaffolds for culturing human prostate cancer (PCa) cells and studying tumor cell interaction with human peripheral blood lymphocytes (PBLs) ex vivo. CA scaffolds and Matrigel matrix samples support in vitro tumor spheroid formation over 15 d of culture, and CA scaffolds support live-cell fluorescence imaging with confocal microscopy using stably transfected PCa cells for 55 d. PCa cells grown in Matrigel matrix and CA scaffolds for 15 d are co-cultured with PBLs for 2 and 6 d in vitro and evaluated with scanning electron microscopy (SEM), immunohistochemistry (IHC), and flow cytometry. Both the Matrigel matrix and CA scaffolds support interaction of PBLs with PCa tumors, with CA scaffolds providing a more robust platform for subsequent analyses. This study demonstrates the use of 3D natural polymer scaffolds as a tissue culture model for supporting long-term analysis of interaction of prostate cancer tumor cells with immune cells, providing an in vitro platform for rapid immunotherapy development.
authors.affiliation: Department of Materials Science and Engineering, University of Washington, 302L Roberts Hall, Box 352120, Seattle, WA, 98195, USA.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Florczyk
authors.forename: Gang
authors.initials: G
authors.lastname: Liu
authors.forename: Forrest M
authors.initials: FM
authors.lastname: Kievit
authors.forename: Allison M
authors.initials: AM
authors.lastname: Lewis
authors.forename: Jennifer D
authors.initials: JD
authors.lastname: Wu
authors.forename: Miqin
authors.initials: M
authors.lastname: Zhang
mesh_terms: D002450:Cell Communication; D045744:Cell Line, Tumor; D018920:Coculture Techniques / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D008214:Lymphocytes / Q000473:pathology* / Q000502:physiology*; D008297:Male; D016062:Porosity; D011471:Prostatic Neoplasms / Q000473:pathology* / Q000503:physiopathology*; D054457:Tissue Scaffolds