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ABSTRACT: Recent studies have suggested that prostaglandin (PG) E2 (PGE2) and the prostaglandin pathway are essential for hematopoietic stem cell growth and development. However, similar studies on hematopoietic commitment from human embryonic stem cells (hESCs) are still limited. Here we report that the addition of PGE2 promotes hematopoietic differentiation of hESCs. The induced cells from hESCs/OP9 co-culture and in the presence of PGE2 were characterized by reverse transcription-PCR (RT-PCR), flow cytometry, colony-forming assays and Wright-Giemsa staining. Our results demonstrated that PGE2 exposure could alter the gene expression pattern and morphology of co-cultured hESCs and resulted in a robust hematopoietic differentiation with higher frequencies of CD34+ and CD45+ cells. Furthermore, the Smad signaling pathway may be involved in PGE2 and OP9 induced hematopoietic differentiation of hESCs. This research may improve our knowledge of stem cell regulation and hopefully lead to better stem cell-based therapeutic options. Keywordshuman embryonic stem cells-prostaglandin E2-hematopoiesis- in vitro differentiation01/2010; 5(5):445-454.