Directed Differentiation of Embryonic Stem Cells Using a Bead-Based Combinatorial Screening Method
Figure 5
Combinatorial screening discovers small molecule-driven differentiation protocols for the generation of lymphoid progenitor cells.
(a) Histogram showing efficiency of the most frequently occurring protocols derived from the CombiCult screen, as well as novel combinations of frequently occurring chemicals, compared to the most efficient cytokine-driven protocol (0185). Phagocytosis activity was measured by pHrodo assay. Mix A: metformin, sis3, kenpaullone and GPR-40 agonist on day 5, followed by pifithrin, capsaicin and DITPA on day 7. Mix B: metformin, sis3, kenpaullone, GPR-40 agonist on day 5, followed by pifithrin, capsaicin, DITPA, transRA, and VEGF inducer on day 7. Mix C: metformin, sis3, kenpaullone, GPR-40 agonist, TEA on day 5, followed by pifithrin, capsaicin, DITPA, transRA, and VEGF inducer on day 7. Each bar represents the average efficiency from 2 wells (each one containing 4000 beads) and the graph is representative of 3 separate experiments. There is a statistically significant difference between the cytokine protocol and the individual mixes *p<0.05. (b) Representative images of lymphoid progenitor cells generated on beads and treated with pHrodo particles (red fluorescence): (i) cytokine-driven protocol 0185 and (ii) chemically-driven protocol (Mix B).