American Society of Hematology

Fig. 5.

$Fig. 5. Hemogenic ability of GFP− or GFP+ endothelial cells. / (A) GFP− or GFP+ endothelial cells (VE cadherin+, CD45−, and Ter119−) were harvested from 2.5-day culture of sorted Flk-1+ cells and cultured on OP9 cells. Then the expression of hematopoietic markers and GFP were analyzed by flow cytometry 24 hours later. The result shown is a representative of 3 independent experiments. (B) May-Grünwald-Giemsa staining of hematopoietic cells formed in the culture of GFP− endothelial cells. Erythroblasts, monocytes, macrophages, and polymorphonuclear cells were observed. The scale bar represents 10 μm. (C) Morphology of a PECAM-1+colony generated from either GFP− or GFP+endothelial cells. The scale bar represents 200 μm. (D) Frequency of cells capable of formation of endothelial colony in the indicated fractions. Error bars indicate standard deviations for 3 independent determinations. (E) Frequency of hematopoietic colony-forming cells in the indicated fractions. Error bars indicate standard deviations for 3 independent determinations. (F) Incidence of hematopoietic and endothelial cell differentiation from single GFP− or GFP+ endothelial cells (VE cadherin+, CD45−, and Ter119−) on OP9 stromal cell layer in the presence of SCF, IL-3, Epo, and G-CSF. Arrows indicate that no colony-forming cell was detected. Error bars indicate standard deviations for 3 independent determinations.$

Hemogenic ability of GFP⁻ or GFP⁺ endothelial cells.

(A) GFP⁻ or GFP⁺ endothelial cells (VE cadherin⁺, CD45⁻, and Ter119⁻) were harvested from 2.5-day culture of sorted Flk-1⁺ cells and cultured on OP9 cells. Then the expression of hematopoietic markers and GFP were analyzed by flow cytometry 24 hours later. The result shown is a representative of 3 independent experiments. (B) May-Grünwald-Giemsa staining of hematopoietic cells formed in the culture of GFP⁻ endothelial cells. Erythroblasts, monocytes, macrophages, and polymorphonuclear cells were observed. The scale bar represents 10 μm. (C) Morphology of a PECAM-1⁺colony generated from either GFP⁻ or GFP⁺endothelial cells. The scale bar represents 200 μm. (D) Frequency of cells capable of formation of endothelial colony in the indicated fractions. Error bars indicate standard deviations for 3 independent determinations. (E) Frequency of hematopoietic colony-forming cells in the indicated fractions. Error bars indicate standard deviations for 3 independent determinations. (F) Incidence of hematopoietic and endothelial cell differentiation from single GFP⁻ or GFP⁺ endothelial cells (VE cadherin⁺, CD45⁻, and Ter119⁻) on OP9 stromal cell layer in the presence of SCF, IL-3, Epo, and G-CSF. Arrows indicate that no colony-forming cell was detected. Error bars indicate standard deviations for 3 independent determinations.

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