title: Controlled Generation of Microspheres Incorporating Extracellular Matrix Fibrils for Three-Dimensional Cell Culture.
abstract: A growing body of evidence suggests that studying cell biology in classical two-dimensional formats, such as cell culture plasticware, results in misleading, non-physiological findings. For example, some aspects of cancer biology cannot be observed in 2D, but require 3D culture methods to recapitulate observations in vivo. Therefore, we developed a microsphere-based model to permit 3D cell culture incorporating physiological extracellular matrix components. Bio-electrospraying was chosen as it is the most advanced method to produce microspheres, with THP-1 cells as a model cell line. Bio-electrospraying parameters, such as nozzle size, polymer flow rate, and voltage, were systematically optimized to allow stable production of size controlled microspheres containing extracellular matrix material and human cells. We investigated the effect of bio-electrospraying parameters, alginate type and cell concentration on cell viability using trypan blue and propidium iodide staining. Bio-electrospraying had no effect on cell viability nor the ability of cells to proliferate. Cell viability was similarly minimally affected by encapsulation in all types of alginate tested (MVM, MVG, chemical- and food-grade). Cell density of 5 × 106cells ml-1within microspheres was the optimum for cell survival and proliferation. The stable generation of microspheres incorporating cells and extracellular matrix for use in a 3D cell culture will benefit study of many diverse diseases and permit investigation of cellular biology within a 3D matrix.
authors.affiliation: BioPhysics Group, UCL Institute of Biomedical Engineering, UCL Centre for Stem Cells and Regenerative Medicine and Department of Mechanical Engineering, University College London, Torrington Place, London, WC1E 7JE, United Kingdom.
authors.forename: Victoria L
authors.initials: VL
authors.lastname: Workman
authors.affiliation: Clinical and Experimental Sciences, Institute for Life Sciences, Faculty of Medicine, University of Southampton, Southampton, SO16 6YD, United Kingdom.
authors.forename: Liku B
authors.initials: LB
authors.lastname: Tezera
authors.affiliation: Clinical and Experimental Sciences, Institute for Life Sciences, Faculty of Medicine, University of Southampton, Southampton, SO16 6YD, United Kingdom.
authors.forename: Paul T
authors.initials: PT
authors.lastname: Elkington
authors.affiliation: BioPhysics Group, UCL Institute of Biomedical Engineering, UCL Centre for Stem Cells and Regenerative Medicine and Department of Mechanical Engineering, University College London, Torrington Place, London, WC1E 7JE, United Kingdom.
authors.forename: Suwan N
authors.initials: SN
authors.lastname: Jayasinghe
keywords: 3D cell culture; bio-electrospraying; biological models; cell encapsulation; cellular kinetics