In order to differentiate T cells in vitro, co-culture systems with Notch ligand-expressing feeder cells have been in use for a long time. Here we describe a feeder-free culture condition for differentiating T cells from hematopoietic cells that are cultured on Fc-DLL4-coated plate with T-lineage cytokines. This condition is capable of efficiently differentiating hematopoietic progenitor cells (HPCs) to immature T cells expressing both CD4 and CD8. To mature those cells into functional T cells, further stimulation and culture is necessary.