Figure 1.

cAMP Induction Increases HSC-like Frequency during hPSC-to-Hematopoietic Differentiation

(A) Conditions and timeline applied to differentiate hPSCs toward mesoderm commitment and hematopoietic differentiation.

(B) Flow cytometric analysis of hematopoietic cells at day 14 of differentiation. Representative flow cytometry plots (biexponential axis) of cells cultured in control medium (MesoTotal), and cells treated with forskolin, IBMX, or forskolin + IBMX are shown.

(C) Percentage of the hematopoietic surface phenotypes indicated in (B). Data represent mean ± SEM of three independent experiments. Statistical analysis was performed using the t test. Significance compared with the control setting: ^∗p < 0.05, ^∗∗∗p < 0.001; n.s., not significant.

(D) Frequency of the putative HSC-like cells (in viable fraction). Data represent mean ± SEM of three independent experiments. Statistical analysis was performed using the t test. Significance compared with the control setting: ^∗p < 0.05; n.s., not significant.

(E) Total CFU numbers after 12-day CFU assay of differentiated hematopoietic cells treated with forskolin, IBMX, and forskolin + IBMX. The CFU distribution of three independent experiments is shown as mean ± SEM CFU-G (granulocyte), CFU-M (macrophage), CFU-E (erythroid), CFU-GM (granulocyte/macrophage). Statistical analysis was performed using the t test. Significance compared with the control setting: ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001.

(F) Cell numbers obtained per well (per 1 × 10⁴ seeded cells) after CFU assay. Data represent mean ± SEM of three independent experiments. Statistical analysis was performed using the t test. Significance compared with the control setting: ^∗∗p < 0.01, ^∗∗∗p < 0.001; n.s., not significant.

See also Figure S1.