Isolation, Culture and Comprehensive Characterization of Biological Properties of Human Urine-Derived Stem Cells

Int J Mol Sci. 2021 Nov 19;22(22):12503. doi: 10.3390/ijms222212503.

Abstract

Mesenchymal stem cells (MSCs) represent an attractive source within the field of tissue engineering. However, their harvesting often requires invasive medical procedures. Urine-derived stem cells (UDSCs) display similar properties to MSCs, and their obtention and further processing is non-invasive for the donors as well as low cost. Here, we offer a comprehensive analysis of their biological properties. The goal of this study was to analyze their morphology, stemness, differentiation potential and cytokine profile. We have successfully isolated UDSCs from 25 urine samples. First colonies emerged up to 9 days after the initial seeding. Cell doubling time was 45 ± 0.24 SD, and when seeded at the density of 100 cells/cm2, they formed 42 ± 6.5 SD colonies within 10 days. Morphological analyzes revealed that two different types of the cell populations have been present. The first type had a rice-grain shape and the second one was characterized by a polyhedral shape. In several cell cultures, dome-shaped cells were observed as well. All examined UDSCs expressed typical MSC-like surface markers, CD73, CD90 and CD105. Moreover, conditioned media from UDSCs were harvested, and cytokine profile has been evaluated showing a significantly higher secretory rate of IL-8, IL-6 and chemokines MCP-1 and GM-CSF. We have also successfully induced human UDSCs into chondrogenic, osteogenic and myogenic cell lineages. Our findings indicate that UDSCs might have immense potential in the regeneration of the damaged tissues.

Keywords: biological characteristics; cytokine profile; differentiation capacity; morphology; stemness; urine-derived stem cells.

MeSH terms

  • Adipogenesis / genetics
  • Cell Culture Techniques
  • Cell Differentiation / genetics*
  • Cell Lineage / genetics
  • Cell Proliferation / genetics*
  • Chemokine CCL2 / genetics
  • Chondrogenesis / genetics
  • Gene Expression Regulation, Developmental / genetics
  • Granulocyte-Macrophage Colony-Stimulating Factor / genetics
  • Humans
  • Interleukin-6 / genetics
  • Interleukin-8 / genetics
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / metabolism
  • Osteogenesis / genetics
  • Stem Cells / cytology*
  • Stem Cells / metabolism

Substances

  • CCL2 protein, human
  • Chemokine CCL2
  • Interleukin-6
  • Interleukin-8
  • Granulocyte-Macrophage Colony-Stimulating Factor