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

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.

High Therapeutic and Esthetic Properties of Extracellular Vesicles Produced from the Stem Cells and Their Spheroids Cultured from Ocular Surgery-Derived Waste Orbicularis Oculi Muscle Tissues

“…The tissues were chopped and the cells from the tissues were cultured in α-MEM supplemented with 10% FBS and 1% PS. The isolation and characterization of AD-MSCs and USCs were carried out as described in our recent publication [ 48 ].…”

Rocking the Boat: The Decisive Roles of Rho Kinases During Oocyte, Blastocyst, and Stem Cell Development

“… Joo et al (2012) made a similar observation during the differentiation and proliferation of endothelial cells derived from an embryonic stem cell (ESC)-derived Flk1+ mesodermal precursor cells. Recently, Kim et al (2020) found that ROCK inhibitor Y-27632 together with Matrigel improved the isolation, proliferation rate, and differentiation potential of urine-derived stem cells in humans. Furthermore, ROCK inhibitors have been used to promote proliferation, viability, and differentiation of mesenchymal stem cells ( Baharvand et al, 2010 ; Mellott et al, 2014 ; Nakamura et al, 2014 ; Li et al, 2015 ).…”

Urine-derived stem cells: applications in skin, bone and articular cartilage repair

“…However, whether USCs cultured by this method remain the same or whether the differences between USCs cultured by different methods lead to different therapeutic outcomes is still unknown. Another solution that faces the same dilemma is reconstituting the culture conditions of the USCs by adding certain nutritional supplements, seeding the USCs on other extracellular matrix components, and taking oxygen tension into account to improve the isolation and proliferation of the USCs [ 67 , 113 ]. Considering batch-to-batch variation, the quality control of the USCs is of great significance.…”