American Society of Hematology

Fig. 3.

$Fig. 3. Hematopoietic potential in Flk1+ fractions from ES-cell/OP9 cocultures. / (A) Day-5 ES-cell/OP9 cocultures were analyzed for Flk1 surface expression (solid line). Staining with control monoclonal antibody (dotted line) is also shown. (B) Flk1+-gated cells were analyzed for the surface expression of CD105 and CD31. (C) Flk1− and Flk1+ cells were isolated from day-5 ES-cell/OP9 cocultures. Flk1+ cells were further fractionated into CD105+CD31+, CD105+CD31−, and CD105−CD31− subsets. An equal number of cells were sorted from each subset, reseeded onto OP9 cells, and analyzed by flow cytometry on subsequent days (with day 0 corresponding to the start of the coculture). The CD45−CD105−quadrants include OP9 stromal cells (d6-d11), and erythrocytes (d11). The total number of cells obtained from each coculture is indicated in parentheses.$

Hematopoietic potential in Flk1⁺ fractions from ES-cell/OP9 cocultures.

(A) Day-5 ES-cell/OP9 cocultures were analyzed for Flk1 surface expression (solid line). Staining with control monoclonal antibody (dotted line) is also shown. (B) Flk1⁺-gated cells were analyzed for the surface expression of CD105 and CD31. (C) Flk1⁻ and Flk1⁺ cells were isolated from day-5 ES-cell/OP9 cocultures. Flk1⁺ cells were further fractionated into CD105⁺CD31⁺, CD105⁺CD31⁻, and CD105⁻CD31⁻ subsets. An equal number of cells were sorted from each subset, reseeded onto OP9 cells, and analyzed by flow cytometry on subsequent days (with day 0 corresponding to the start of the coculture). The CD45⁻CD105⁻quadrants include OP9 stromal cells (d6-d11), and erythrocytes (d11). The total number of cells obtained from each coculture is indicated in parentheses.

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