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An in vitro model of CD34 + CD38 −  stem cell (SC) differentiation in postnatal cultured thymic epithelia fragment (CTEF) cocultures is described. Sequential phenotypic analysis of the progeny of the SC‐CTEF demonstrated predominantly thymocytes and minor populations of promyelocytes, monocytes and natural killer cells. Triple‐positive CD3 + CD4 + CD8 + , double‐positive CD4 + CD8 + , and mature single‐positive CD4 +  and CD8 +  T cells, which were TCRαβ + , were identified indicating normal thymocyte maturation. In kinetic studies, mature single‐positive CD4 +  T cells increased from 29% of total cells at one week to 54% at four weeks of coculture. These findings demonstrate that coculture of bone marrow‐derived SC and allogeneic cultured thymic epithelia in vitro results in continuous normal predominantly thymocyte differentiation. The SC‐CTEF cocultures were then infected with two different strains of human immunodeficiency virus. CD4 +  thymocytes were markedly decreased. However, inhibition of early thymocyte maturation steps was also suggested by the presence of increased triple‐negative and CD44 + CD25 − CD3 −  thymocytes and decreased CD44 + CD25 +  thymocytes. This model system of thymocyte maturation will be useful in the evaluation of primary T cell immunodeficiency disorders, gene therapy of SC and pharmacological augmentation of thymic function.

stem cells

Development of a Method of Thymocyte Differentiation of Bone Marrow‐Enriched CD34 + CD38 −  Cells in Postnatal Allogeneic Cultured Thymic Epithelia to Evaluate Immunodeficiency Disorders