Thrombocytopenia is a common problem in hematology/oncology patients. In the past two decades a number of thrombopoietic growth factors and related cytokines have become available for clinical investigations. Unfortunately, most of the pleiotropic cytokines have been limited by their modest activity and toxicity profile. The discovery of thrombopoietin (TPO), a key regulator of platelet production, led to the clinical development of two recombinant versions of the molecule: full-length, recombinant human thrombopoietin (rhTPO), and truncated and pegylated, megakaryocyte growth and development factor (Peg-rHuMGDF). Both agents showed significant biologic activity in various clinical settings, including nonmyeloablative chemotherapy, mobilization of progenitors, platelet apheresis, and treatment of thrombocytopenia related to other conditions. Despite promising thrombopoietic activity, the clinical development of the first generation of recombinant TPOs was discontinued due to the neutralizing antibodies observed with PEG-rHuMGDF. This has led to the development of TPO agonists with no sequence homology to TPO, which can bind to the TPO receptors and activate signaling, leading to an increase in platelet production. The clinical experience with the first generation of thrombopoietic agents has provided insight into the biology and future directions for a second generation of thrombopoietic agents in various disorders of thrombocytopenia.