J Clin Med. 2020 Mar; 9(3): 827. Published online 2020 Mar 18. doi: 10.3390/jcm9030827

Improved Isolation and Culture of Urine-Derived Stem Cells (USCs) and Enhanced Production of Immune Cells from the USC-Derived Induced Pluripotent Stem Cells

Kyeongseok Kim,^1,† Minchan Gil,^1,† Ahmed Abdal Dayem,¹ Sangbaek Choi,¹ Geun-Ho Kang,¹ Gwang-Mo Yang,¹ Sungha Cho,¹ Yeojin Jeong,¹ Se Jong Kim,¹ Jaekwon Seok,¹ Hee Jeong Kwak,¹ Subbroto Kumar Saha,¹ Aram Kim,² and Ssang-Goo Cho^1,*

Abstract

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.