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. 2010 Oct 5;21(11):1555–1567. doi: 10.1089/hum.2010.050

FIG. 4.

FIG. 4.

hiPSC clones derived from HFFs by infection of the FRh11 encoding human reprogramming factors share hESC character. (A) Representative images of hESC and hiPSC #11 colonies plated on Matrigel. (B) Immunostaining of hESC and hiPSC #11 colonies for Nanog. hESCs and hiPSC #11 were fixed with 1.0% formaldehyde/PBS and permeabilized with 0.2% Triton X-100/PBS. The cells were then stained with a rabbit anti-Nanog antibody and a DyLight 488 anti-rabbit IgG. 7-AAD was used for nuclear staining. (C) Cell-surface expression of hESC markers on hiPSC #11. hESCs and hiPSC #11 were dissociated into single-cell suspensions with a trypsin-EDTA and stained with specific antibodies against SSEA1, SSEA3, SSEA4, TRA-1-60, and TRA-1-81. The cells were then analyzed by flow cytometry. Numbers (%) in each population are shown on each plot. (D) Dark-field images of EBs generated from hESCs and hiPSC #11. hESCs and hiPSC #11 were dissociated into single-cell suspensions with Accutase. The size-controlled EBs (3,000 cells/EB) were formed with an AggreWell 400 plate and maintained in StemLine II medium for 4 days. Color images available online at www.liebertonline.com/hum.