Self-Renewal and Differentiation Capacity of Urine-Derived Stem Cells after Urine Preservation for 24 Hours

Abstract

Despite successful approaches to preserve organs, tissues, and isolated cells, the maintenance of stem cell viability and function in body fluids during storage for cell distribution and transportation remains unexplored. The aim of this study was to characterize urine-derived stem cells (USCs) after optimal preservation of urine specimens for up to 24 hours. A total of 415 urine specimens were collected from 12 healthy men (age range 20–54 years old). About 6×104 cells shed off from the urinary tract system in 24 hours. At least 100 USC clones were obtained from the stored urine specimens after 24 hours and maintained similar biological features to fresh USCs. The stored USCs had a “rice grain” shape in primary culture, and expressed mesenchymal stem cell surface markers, high telomerase activity, and normal karyotypes. Importantly, the preserved cells retained bipotent differentiation capacity. Differentiated USCs expressed myogenic specific proteins and contractile function when exposed to myogenic differentiation medium, and they expressed urothelial cell-specific markers and barrier function when exposed to urothelial differentiation medium. These data demonstrated that up to 75% of fresh USCs can be safely persevered in urine for 24 hours and that these cells stored in urine retain their original stem cell properties, indicating that preserved USCs could be available for potential use in cell-based therapy or clinical diagnosis.

Silver nanoparticles promote osteogenic differentiation of human urine-derived stem cells at noncytotoxic concentrations

“…Four 250-300 mL mid stream urine samples from four healthy men aged 28-34 years were collected and directly transferred to the laboratory for isolation and culture, as reported previously, [2][3][4][5]7,8 with a slight modification. Briefly, urine specimens were centrifuged at 400× g for 10 minutes at room temperature.…”

“…Qin et al from urine and expanded in special culture, can differentiate into cell lineages such as urothelial cells, smooth muscle cells, endothelial cells, osteocytes, chondrocytes, and adipocytes on induction with appropriate media, [4][5][6][7][8] and have been reported recently to be ideal candidates for mesenchymal stem cells to replace marrow-derived stem cells and adipose-derived stem cells as seed cells for tissue engineering, due to their harvest from autologous urine using noninvasive, safe, low-cost, and easily reproducible methods. 3,7 Currently, USCs are being investigated as cell therapies in the treatment of renal insufficiency and urinary incontinence, and for tissue engineering in bladder and urethral tissue regeneration; 6 however, their use in tissue-engineering to repair bone defects is equally attractive.…”

“…USCs at passage 3 were characterized using flow cytometry analysis as described elsewhere. [3][4][5] Briefly, cells were trypsinized, washed in ice-cold phosphate-buffered saline containing 0.5% bovine serum albumin (wash buffer), and counted. Fluorochrome-conjugated antibodies, anti-CD34-fluorescein isothiocyanate (FITC), CD45-FITC, CD90-FITC, CD29-phycoerythrin, and CD73-phycoerythrin (BD Pharmingen, Franklin Lakes, NJ, USA), were added to 5 × 10 5 cells in 50 µL of phosphate-buffered saline containing 3% bovine serum albumin and incubated on ice for 30 minutes in the dark.…”

Isolation and in vitro cultivation of human urine-derived cells: an alternative stem cell source

“…To overcome this limitation, studies have especially focused on autologous stem cells derived from a variety of adult tissues such as muscle, bone marrow and adipose tissue. [2] Moreover, it has recently been demonstrated that autologous stem cells can also be obtained from urine by a noninvasive and low-cost technique. [3,4] Therefore, urine can be an alternative autologous stem cell source for cell-based therapies.…”

“…For example, urine preservation conditions which are suitable for retaining proliferation and multipotent differentiation capabilities of stem cells in fresh urine samples have been explained in only one paper. [2] Additionally, little data are available about age, gender and health status of urine donors. [1,2,[4][5][6] Therefore, development of a reliable method for preservation of cells in urine will increase the amount of high quality cells obtained, and also will alleviate cell damage caused by storing them in urine.…”