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. 2017 Oct 19;9(5):1369–1376. doi: 10.1016/j.stemcr.2017.09.016

Figure 1.

Figure 1

Differentiating Healthy Control hiPSCs into Osteoclasts

(A) Schematic protocol of differentiating hiPSCs into osteoclasts.

(B) Embryoid body (EB) formation and mesoderm gene expression. EBs cultured for 4 days (left panel). Scale bar, 200 μm. Expression of mesoderm marker genes in EBs cultured for 1, 2, 3, and 4 days by qPCR. ∗p < 0.05 by one-way ANOVA. Data presented are means ± SD.

(C) Myelomonocytic cell expansion. Single cells released from EBs into suspension (top panel). Scale bar, 100 μm. Percentage of cells positive for hematopoietic cell surface markers CD14, CD43, and CD45 in cells released from 10, 13, 17, and 21 day adherent EBs by flow cytometry. Data presented are means ± SD.

(D) TRAP+ osteoclasts differentiated from hiPSCs (left panel), resorption pits on bone chips (middle panel), and expression of OC marker genes, TRAP and CTSK by RT-PCR (right panel). HPRT served as internal control. Ctl1, control1; Ctl2, control2; 1w, 2w, 1 or 2 weeks in stage 3. Scale bar, 100 μm (left panel) and 200 μm (middle panel). Three independent experiments (three technical replicates per experiment) for each hiPSC line. Data were pooled from four wild-type hiPSC lines (hiPSCs from two healthy subjects, two hiPSC clones of each individual donor).