Diverse T-cell differentiation potentials of human fetal thymus, fetal liver, cord blood and adult bone marrow CD34 cells on lentiviral Delta-like-1-modified mouse stromal cells.
Human haematopoietic progenitor/stem cells (HPCs) differentiate into functional T cells in the thymus through a series of checkpoints. A convenient in vitro system will greatly facilitate the understanding of T-cell development and future engineering of therapeutic T cells. In this report, we established a lentiviral vector-engineered stromal cell line (LSC) expressing the key lymphopoiesis regulator Notch ligand, Delta-like 1 (DL1), as feeder cells (LSC-mDL1) supplemented with Flt3 ligand (fms-like tyrosine kinase 3, Flt3L or FL) and interleukin-7 for the development of T cells from CD34(+) HPCs. We demonstrated T-cell development from human HPCs with various origins including fetal thymus (FT), fetal liver (FL), cord blood (CB) and adult bone marrow (BM). The CD34(+) HPCs from FT, FL and adult BM expanded more than 100-fold before reaching the beta-selection and CD4/CD8 double-positive T-cell stage. The CB HPCs, on the other hand, expanded more than 1000-fold before beta-selection. Furthermore, the time required to reach beta-selection differed for the various HPCs, 7 days for FT, 14 days for FL and CB, and 35 days for adult BM. Nevertheless, all of the T cells developed in vitro were stalled at the double-positive or immature single-positive stage with the exception that some CB-derived T cells arrived at a positive selection stage. Consequently, the LSC-mDL1 culture system illustrated diverse T-cell development potentials of pre- and post-natal and adult human BM HPCs. However, further modification of this in vitro T-cell development system is necessary to attain fully functional T cells.
Patel
Ekta
E
Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida, Gainesville, FL 32610-0266, USA.
Wang
Bei
B
Lien
Lily
L
Wang
Yichen
Y
Yang
Li-Jun
LJ
Moreb
Jan S
JS
Chang
Lung-Ji
LJ
D000818:Animals; D001854:Bone Marrow Cells / Q000187:drug effects / Q000502:physiology; D002135:Calcium-Binding Proteins; D002454:Cell Differentiation; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D005312:Fetal Blood / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D005333:Fetus / Q000166:cytology; D006412:Hematopoietic Stem Cells / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D015851:Interleukin-7 / Q000494:pharmacology; D016086:Lentivirus; D008099:Liver / Q000166:cytology / Q000196:embryology; D008565:Membrane Proteins / Q000494:pharmacology; D051379:Mice; D017154:Stromal Cells / Q000187:drug effects / Q000502:physiology; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D013950:Thymus Gland / Q000166:cytology / Q000196:embryology / Q000276:immunology*; D014161:Transduction, Genetic
D002135:Calcium-Binding Proteins; C512925:Dlk1 protein, mouse; D036341:Intercellular Signaling Peptides and Proteins; D015851:Interleukin-7; D008565:Membrane Proteins; C084881:flt3 ligand protein