Tissue factor differential expression in trophoblasts and hematopoietic cells derived from human embryonic stem cells. (A) Schematic diagram of human embryonic stem cell (hESC) differentiation into hematopoietic and trophoblastic cells. (B) Immunostaining showed hESCs expressing Nanog. (C) Trophoblastic differentiation of H9 cells. H9 hESCs were induced in medium with BMP4 to differentiate into trophoblasts. Expression of Oct-4, Nanog, CDX2, and tissue factor (TF) were examined using reverse-transcription polymerase chain reaction (PCR). (D) H9 hESCs were induced to differentiate into hematopoietic cells. Florescence-activated flow cytometry (FACS) analysis showed that TF (CD142) was expressed in CD14+ and CD15+ G-M cells, but not in CD34+ hematopoietic stem/progenitor cells (HSPCs) and CD235+ erythrocytes. (E) TF mRNA in different types of hematopoietic cells was examined by quantitative real-time PCR. (F) TF protein in different types of hematopoetic cells and trophoblasts was examined by western blotting. APC, allophycocyanin; bp, base pair; FITC, fluorescein isothiocyanate; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; G-M, granulocyte–macrophage; PE, phycoerythrin.