Progenitor T-cell differentiation from hematopoietic stem cells using Delta-like-4 and VCAM-1

Nat Methods. 2017 May;14(5):531-538. doi: 10.1038/nmeth.4258. Epub 2017 Apr 10.


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.

MeSH terms

  • Biotechnology / methods
  • CD3 Complex / immunology
  • Cell Differentiation*
  • Hematopoietic Stem Cells / cytology*
  • Hematopoietic Stem Cells / immunology
  • Humans
  • Intracellular Signaling Peptides and Proteins / metabolism*
  • Membrane Proteins / metabolism*
  • Signal Transduction
  • T-Lymphocytes / cytology*
  • T-Lymphocytes / immunology
  • Thymus Gland / cytology
  • Thymus Gland / immunology
  • Vascular Cell Adhesion Molecule-1 / metabolism*


  • CD3 Complex
  • Intracellular Signaling Peptides and Proteins
  • Membrane Proteins
  • Vascular Cell Adhesion Molecule-1
  • delta protein