title: Progenitor T-cell differentiation from hematopoietic stem cells using Delta-like-4 and VCAM-1.
abstract: The molecular and cellular signals that guide T-cell development from hematopoietic stem and progenitor cells (HSPCs) remain poorly understood. The thymic microenvironment integrates multiple niche molecules to potentiate T-cell development in vivo. Recapitulating these signals in vitro in a stromal cell-free system has been challenging and limits T-cell generation technologies. Here, we describe a fully defined engineered in vitro niche capable of guiding T-lineage development from HSPCs. Synergistic interactions between Notch ligand Delta-like 4 and vascular cell adhesion molecule 1 (VCAM-1) were leveraged to enhance Notch signaling and progenitor T-cell differentiation rates. The engineered thymus-like niche enables in vitro production of mouse Sca-1+cKit+and human CD34+HSPC-derived CD7+progenitor T-cells capable of in vivo thymus colonization and maturation into cytokine-producing CD3+T-cells. This engineered thymic-like niche provides a platform for in vitro analysis of human T-cell development as well as clinical-scale cell production for future development of immunotherapeutic applications.
authors.affiliation: Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Shreya
authors.initials: S
authors.lastname: Shukla
authors.affiliation: Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Langley
authors.affiliation: Department of Immunology, Sunnybrook Research Institute, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Jastaranpreet
authors.initials: J
authors.lastname: Singh
authors.affiliation: Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada.
authors.forename: John M
authors.initials: JM
authors.lastname: Edgar
authors.affiliation: Department of Immunology, Sunnybrook Research Institute, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Mahmood
authors.initials: M
authors.lastname: Mohtashami
authors.affiliation: Department of Immunology, Sunnybrook Research Institute, University of Toronto, Toronto, Ontario, Canada.|Medicine by Design, a Canada First Research Excellence Program at the University of Toronto, Toronto, Ontario, Canada.
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Zúñiga-Pflücker
authors.affiliation: Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada.|Medicine by Design, a Canada First Research Excellence Program at the University of Toronto, Toronto, Ontario, Canada.|Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada.|Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Peter W
authors.initials: PW
authors.lastname: Zandstra
mesh_terms: D001709:Biotechnology / Q000379:methods; D017252:CD3 Complex / Q000276:immunology; D002454:Cell Differentiation; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000276:immunology; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000166:cytology* / Q000276:immunology; D013950:Thymus Gland / Q000166:cytology / Q000276:immunology; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism*
chemical_list: D017252:CD3 Complex; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D019010:Vascular Cell Adhesion Molecule-1; C064045:delta protein