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. Author manuscript; available in PMC: 2017 Mar 7.
Published in final edited form as: Dev Cell. 2016 Mar 7;36(5):525–539. doi: 10.1016/j.devcel.2016.02.011

Figure 1. Prominin1, Sca1 and CD34 Mark HP Cells in Midgestation Mouse Placentas.

Figure 1

(A) Strategy to isolate HP cells in vivo using information generated from in vitro hemogenic induction.

(B) E10.5 and E12.5 placentas were isolated and dissociated to a single cell suspension and (C) analyzed for expression of Prom1, Sca1, CD34 and CD45, red boxes highlight populations of interest.

(D) Percentage of Prom1+Sca1+CD34+ cells (PS34, left panel) as well as the percentage of the subpopulation of PS34CD45− cells (right panel) from litters at E10.5, E11.5 and E12.5 (each mark represents a single placenta, n=11–28). *p<0.05; ***p<0.001.

(E) PS34CD45− cells were sorted and plated on gelatin-coated dishes in Myelocult with SCF, IL-3 and Flt3l for 7 days, stained for CD45 and analyzed by flow cytometry or (F) immunofluorescence. Dashed lines highlight large adherent cells associated with round non-adherent CD45+ cells.

(G) PS34CD45− cells were cultured on AFT024 stroma for 4 weeks; cobblestone areas are stained for CD45. Scale bar = 100 μm.