title: Programmable 3D silk bone marrow niche for platelet generation ex vivo and modeling of megakaryopoiesis pathologies.
abstract: We present a programmable bioengineered 3-dimensional silk-based bone marrow niche tissue system that successfully mimics the physiology of human bone marrow environment allowing us to manufacture functional human platelets ex vivo. Using stem/progenitor cells, megakaryocyte function and platelet generation were recorded in response to variations in extracellular matrix components, surface topography, stiffness, coculture with endothelial cells, and shear forces. Millions of human platelets were produced and showed to be functional based on multiple activation tests. Using adult hematopoietic progenitor cells our system demonstrated the ability to reproduce key steps of thrombopoiesis, including alterations observed in diseased states. A critical feature of the system is the use of natural silk protein biomaterial allowing us to leverage its biocompatibility, nonthrombogenic features, programmable mechanical properties, and surface binding of cytokines, extracellular matrix components, and endothelial-derived proteins. This in turn offers new opportunities for the study of blood component production ex vivo and provides a superior tissue system for the study of pathologic mechanisms of human platelet production.
authors.affiliation: Department of Molecular Medicine, University of Pavia, Pavia, Italy; Biotechnology Research Laboratories, Istituto di Ricovero e Cura a Carattere Scientifico San Matteo Foundation, Pavia, Italy;
authors.forename: Christian A
authors.initials: CA
authors.lastname: Di Buduo
authors.affiliation: Department of Molecular Medicine, University of Pavia, Pavia, Italy; Biotechnology Research Laboratories, Istituto di Ricovero e Cura a Carattere Scientifico San Matteo Foundation, Pavia, Italy; Department of Biomedical Engineering, Tufts University, Medford, MA;
authors.forename: Lindsay S
authors.initials: LS
authors.lastname: Wray
authors.affiliation: Department of Molecular Medicine, University of Pavia, Pavia, Italy; Biotechnology Research Laboratories, Istituto di Ricovero e Cura a Carattere Scientifico San Matteo Foundation, Pavia, Italy; Department of Biomedical Engineering, Tufts University, Medford, MA;
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Tozzi
authors.affiliation: Department of Molecular Medicine, University of Pavia, Pavia, Italy; Biotechnology Research Laboratories, Istituto di Ricovero e Cura a Carattere Scientifico San Matteo Foundation, Pavia, Italy;
authors.forename: Alessandro
authors.initials: A
authors.lastname: Malara
authors.affiliation: Department of Biomedical Engineering, Tufts University, Medford, MA;
authors.forename: Ying
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Biomedical Engineering, Tufts University, Medford, MA;
authors.forename: Chiara E
authors.initials: CE
authors.lastname: Ghezzi
authors.affiliation: Department of Biomedical Engineering, Tufts University, Medford, MA;
authors.forename: Daniel
authors.initials: D
authors.lastname: Smoot
authors.affiliation: Department of Biomolecular Sciences, Biochemistry and Molecular Biology Section, University of Urbino "Carlo Bo," Urbino, Italy;
authors.forename: Carla
authors.initials: C
authors.lastname: Sfara
authors.affiliation: Department of Biomolecular Sciences, Biochemistry and Molecular Biology Section, University of Urbino "Carlo Bo," Urbino, Italy;
authors.forename: Antonella
authors.initials: A
authors.lastname: Antonelli
authors.affiliation: Department of Physics, Tufts University, Medford, MA;
authors.forename: Elise
authors.initials: E
authors.lastname: Spedden
authors.affiliation: Department of Chemistry, Physical Chemistry Section, University of Pavia, Pavia, Italy;
authors.forename: Giovanna
authors.initials: G
authors.lastname: Bruni
authors.affiliation: Department of Physics, Tufts University, Medford, MA;
authors.forename: Cristian
authors.initials: C
authors.lastname: Staii
authors.affiliation: Department of Translational Research, Stem Cells Unit, Istituto di Ricovero e Cura a Carattere Scientifico Centro di Riferimento Oncologico, Aviano, Italy; and Department of Molecular and Experimental Research, The Scripps Research Institute, La Jolla, CA.
authors.forename: Luigi
authors.initials: L
authors.lastname: De Marco
authors.affiliation: Department of Biomolecular Sciences, Biochemistry and Molecular Biology Section, University of Urbino "Carlo Bo," Urbino, Italy;
authors.forename: Mauro
authors.initials: M
authors.lastname: Magnani
authors.affiliation: Department of Biomedical Engineering, Tufts University, Medford, MA;
authors.forename: David L
authors.initials: DL
authors.lastname: Kaplan
authors.affiliation: Department of Molecular Medicine, University of Pavia, Pavia, Italy; Biotechnology Research Laboratories, Istituto di Ricovero e Cura a Carattere Scientifico San Matteo Foundation, Pavia, Italy; Department of Biomedical Engineering, Tufts University, Medford, MA;
authors.forename: Alessandra
authors.initials: A
authors.lastname: Balduini
mesh_terms: D000328:Adult; D000818:Animals; D001792:Blood Platelets / Q000166:cytology* / Q000378:metabolism; D012831:Bombyx; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D002478:Cells, Cultured; D018920:Coculture Techniques; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism; D005109:Extracellular Matrix; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D008533:Megakaryocytes / Q000166:cytology* / Q000378:metabolism; D055728:Primary Myelofibrosis / Q000378:metabolism / Q000473:pathology*; D047011:Silk / Q000737:chemistry*; D034061:Thrombopoiesis / Q000502:physiology; D023822:Tissue Engineering; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D047011:Silk