Ecient maintenance of the undierentiated status of human pluripotent stem cells
(hiPSCs) is crucial for producing cells with improved proliferation, survival and dierentiation,
which can be successfully used for stem cell research and therapy. Here, we generated iPSCs
from healthy donor peripheral blood mononuclear cells (PBMCs) and analyzed the proliferation
and dierentiation capacities of the generated iPSCs using single cell NGS-based 24-chromosome
aneuploidy screening and RNA sequencing. In addition, we screened various natural compounds
for molecules that could enhance the proliferation and dierentiation potential of hiPSCs. Among
the tested compounds, 3,20-dihydroxyflavone (3,20-DHF) significantly increased cell proliferation
and expression of naïve stemness markers and decreased the dissociation-induced apoptosis
of hiPSCs. Of note, 3,20-DHF-treated hiPSCs showed upregulation of intracellular glutathione
(GSH) and an increase in the percentage of GSH-high cells in an analysis with a FreSHtracer
system. Interestingly, culture of the 3,20-DHF-treated hiPSCs in dierentiation media enhanced their
mesodermal dierentiation and dierentiation into CD34+ CD45+ hematopoietic progenitor cells
(HPC) and natural killer cells (NK) cells. Taken together, our results demonstrate that the natural
compound 3,20-DHF can improve the proliferation and dierentiation capacities of hiPSCs and
increase the eciency of HPC and NK cell production from hiPSCs.