Hematopoietic potential of the CD45⁻TER119⁻ 4-integrin⁺ cells induced from ES cell-derived FLK1⁺ cells. FLK1⁺ cells sorted from differentiating CCE cells were cultured in a gelatin-coated dish for 3 days. Cultured cells were fractionated by FACS into VE-cadherin⁺ 4-integrin⁻, VE-cadherin⁺ 4-integrin⁺, VE-cadherin⁻ 4-integrin⁻, and VE-cadherin⁻ 4-integrin⁺ cells. CD45⁺ cells and TER119⁺ cells were excluded from the sorting gates. Sorted cells were cultured on OP9 stromal cell layer in the presence of IL-3, Epo, G-CSF, and MGF for 7 days. (A) Reanalyses of the sorted cells. A representative result of more than five independent experiments is shown. (B) Frequency of hematopoietic colony-forming cells in the indicated fractions. The arrow indicates that no colony-forming cell was detected. Error bars indicate standard deviations for four independent determinations. (C) Morphology of a hematopoietic cell colony formed in the culture of VE-cadherin⁺ 4-integrin⁺ cells. Hemoglobinized erythrocytes are observed. The bar represents 100 μm.