This study was aimed to explore the differentiation of in vitro induced human pluripotent stem cells (iPSC) into hematopoietic stem progenitor cells. The human iPSC were induced to differentiate into hematopoietic stem/progenitor cell by co-culturing with OP9 bone marrow stromal cells. The expression of hematopoietic stem/progenitor cell surface markers were detected by flow cytometry. The regulation gene expressions of iPSC and hematopoietic stem/progenitor cells were measured by real-time PCR. The CD34(+) hematopoietic stem/progenitor cells were isolated by using immunomagnetic beads, and were used for colony formation assay. The results showed that after iPSC were co-cultured with OP9 cells for 4 days, the morphological changes of iPSC could be observed. Hematopoietic stem/progenitor cell surface markers CD34 and CD43 could be detected by flow cytometry after differentiation. The pluripotent marker gene OCT4 expression gradually decreased and blood-related transcription factor Gata-2 expression gradually increased, while Runx-1 expression was wavily changed, CD34 expression gradually increased. The erythroid colony(CFU-E), granulocyte colony(CFU-G), megakaryocytic colony(CFU-M), granulocyte-megakaryocytic colony(CFU-GM), and mixed colony(CFU-GEMM) were obtained after cultures for 14 d. It is concluded that the human iPSC cells can be induced to differentiate into hematopoietic stem/progenitor cells in vitro by co-culture with OP9 cells.