The availability of autologous adult stem cells is one of the essential prerequisites for
human stem cell therapy. Urine-derived stem cells (USCs) are considered as desirable cell sources for
cell therapy because donor-specific USCs are easily and non-invasively obtained from urine. Efficient
isolation, expansion, and differentiation methods of USCs are necessary to increase their availability.
Here, we developed a method for efficient isolation and expansion of USCs using Matrigel, and the
rho-associated protein kinase (ROCK) inhibitor, Y-27632. The prepared USCs showed significantly
enhanced migration, colony forming capacity, and differentiation into osteogenic or chondrogenic
lineage. The USCs were successfully reprogramed into induced pluripotent stem cells (USC-iPSCs)
and further differentiated into kidney organoid and hematopoietic progenitor cells (HPCs). Using
flavonoid molecules, the isolation efficiency of USCs and the production of HPCs from the USC-iPSCs
was increased. Taken together, we present an improved isolation method of USCs utilizing Matrigel,
a ROCK inhibitor and flavonoids, and enhanced differentiation of USC-iPSC to HPC by flavonoids.
These novel findings could significantly enhance the use of USCs and USC-iPSCs for stem cell research
and further application in regenerative stem cell-based therapies.