On the basis of reports that deoxyguanosine is selectively toxic for adult T lymphocytes, the usefulness of this compound in the production of lymphocyte‐depleted embryonic thymus rudiments in an in vitro organ culture system was investigated. The results showed that a period of exposure to deoxyguanosine causes depletion of the lymphoid cells while the stromal elements continue to survive, with many of the cells showing an epithelial morphology and expression of I region products of the major histocompatibility complex (MHC). When associated with either fetal liver or another thymus fragment as a source of T cell precursors in transfilter experiments, these “empty” thymuses become recolonized, enabling the production of chimeric thymus with stromal and lymphoid cells of different haplotypes. In combination with functional assays, this system offers an entirely in vitro approach to questions relating to the repertoire potential of T cell precursors from different sources and the role of the thymus in tolerance and MHC restriction.