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Method of Isolation and Culture of Urine-Derived Stem Cells from Pediatric Patients with Primary Nephrotic Syndrom
Shi Dongmei, Li Hao, Zhang Yuanyuan, Gong Mengjia, Yang Haiping, Yang Baohui, Wang Mo*, Li Qiu*
Department of Nephrology, Children’s Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing Key Laboratory of Child Infection and Immunity, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Chongqing 400014, China
Abstract: The aim of this study was to establish a method for isolation and culture of urine-derived stem cells (USCs) from pediatric patients with primary nephrotic syndrome (PNS) (p-USCs). Cells were separated by centrifugation and adherent culture. The morphology， growth curve and cell cycle of cells were observed. The expression of surface markers of cells was analysed by flow cytometry. Osteogenic and adipogenic differentiation potential of cells were detected by alizarin red and Oil Red-O dye staining. We found that cells were grainlike appearance and logarithmic growth. Cells expressed surface markers associated with pericytes (CD146) and mesenchymal stem cells， such as CD24 (cluster of differentiation 24)， CD29， CD73 and CD90， but a few cells expressed CD105， while didn’t express hematopoietic stem cell surface marker CD34. Both of osteogenicdifferentiated and adipogenic-differentiated cells were positive by alizarin red staining and Oil Red-O staining，respectively. These results showed that the method of isolation and culture USCs from pediatric patients with PNS was established successfully， and provided an initial methods for further investigating the potential application of p-USCs in the diagnosis of early renal injury of PNS patients and the treatment of refractory nephrotic syndrome.
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