Hematopoietic cell lines are often used as representatives for a certain cell differentiation lineage and stage, particularly in immunological and hematological studies. Acute lymphoblastic leukemia (ALL) of T-cell type is a rather heterogeneous group of ALL at least by immunophenotyping. Our aim was to present a comprehensive characterization of frequently used T-cell leukemia cell lines and to suggest a correlation with the normal differentiation pattern. A total of 16 T-ALL cell lines were analyzed for their immunophenotype and for T-cell receptor (TCR) rearrangement and expression. The panel of 20 cell surface markers included two new monoclonal antibodies (MoAb), TC-12 and TH-111, which were raised in our laboratory and detect subpopulations of T-cell ALL. TC-12 was typed 'unique', TH-111 was assigned to the CD96 cluster at the Vth Conference on human leucocyte differentiation antigens (HLDA). We categorized the 16 cell lines into the four groups pro-T, pre-T, cortical T and mature T differentiation stage according to the recent proposal of the European Group for the Immunological Characterization of Leukemias (EGIL). Interestingly, none of the T-cell lines were found to be alike. In conclusion, it appears necessary to consider the particular differentiation stage of each individual cell line when using T-cell leukemia lines as models for malignant or normal T cells.