Isolation and Identification of Urine Derived Stem Cells from Patients with Diabetic Nephropathy and Comparison with Urine Derived Stem Cells from Healthy Peoplein Biological Characteristics

Tao Li1,2, Ma Wenjun1,2, Gong Mengjia2 , Bi Yang2 , Wei Guanghui1 , Zhang Yuanyuan1,2*

1 Department of Urology, Children’s Hospital of Chongqing Medical University, Chongqing 400014, China; 2 Laboratory of Stem Cell Biology and Therapy, Children’s Hospital of Chongqing Medical University, Chongqing Key Laboratory of Children Urogenital Development and Tissue Engineering, Ministry of Education Key Laboratory of Child Development and Disorders, Key Laboratory of Pediatrics in Chongqing, Chongqing International Science and Technology Cooperation Center for Child Development and Disorders, Chongqing 400014, China)

Abstract: Stem cell transplantation is a new therapy for diabetic nephropathy， but there are some problems such as ethical and immune rejection of xenograft. In present study， we extracted a type of cell which was similar to stem cells from the urine of diabetic nephropathy patients， called urinary stem cell from patients with diabetic nephropathy (D-USCs). And we compared the biological characteristics between D-USCs and urine derived stem cells (USCs) of healthy people. The potential differentiation ability of the cells was examined by the analysis of cell morphology and surface markers via flow cytology and the staining of alizarin red and oil red O， respectively. We also mapped the growth curve to compare D-USCs and USCs of healthy people proliferative capacity. Human angiogenic factor assay kit was applied to examine the expression of extracellular secretion factor， performed flow cytology to detect cell apoptosis. Our results showed that the cells could be cultured in vitro continuously， and that their morphology remained the size of the grains. These cells were shown to expressing mesenchymal stem cell markers (including CD 24， CD 29， CD 73， CD 90， CD 105) and weakly cell markers (CD 146) in a lower level. And these cells did not express hematopoietic stem cell markers (including CD 31， CD 34 and CD 45). It also had bone and lipid differentiation potential. Compared with USCs， the proliferative ability of D-USCs was damaged， the secretion factor was basically consistently， the number was decreased and the apoptosis rate was increased. In summary， our results showed that D-USCs were successfully extracted from urine of patients with diabetic nephropathy. Our findings showed that D-USCs might provide a new source of cells for the treatment of renal damage by stem cell transplantation with avoiding allogeneic immune rejection.

CSTR: 32200.14.cjcb.2018.02.0005