Biological characteristics of urine-derived stem cells from paediatrics patients with primary nephrotic syndrome

Abstract:

Objective：To investigate and compare the biological characteristics of urine-derived stem cells（USCs） from paediatrics patients with primary nephrotic syndrom（p-USCs） and normal children. Methods：The clean midstream urine was collected from nor－mal children and paediatrics patients with simple primary nephrotic syndrome. USCs and p-USCs were cultured and their morphology features were observed by optical microscopy. The cell reproductive capacity was detected by CCK-8； apoptosis difference and stem cells’ surface markers were analyzed by flow cytometery（CD24，CD29，CD34，CD73，CD90，CD105，CD146）；kidney markers（Pax2） was detected by immunofluorescence and Western blot；autophagy levels was detected by Western blot. Results：Two types of cells were found in p-USCs：p-USCs Ⅰ and p-USCs Ⅱ. p-USCs Ⅰ had the same morphology with USCs：rice-like，small cell body and closely arranged. p-USCs Ⅱ had bigger cell body，small vacuoles and was dispersedly arranged. The largest passage of USCs，p-USCsⅠ and p-USCs Ⅱ were 11，8 and 5 respectively. Compared with those of USCs，the proliferation of p-USCs was decreased，p-USCs Ⅱ in particular（F=217.300，P=0.000）；no statistic difference in the apoptotic rate and expression of stem cell surface markers show was observed between USCs and p-USCs. Compared with those of USCs，the expression of Pax2 was decreased in p-USCs，but only p-USCs Ⅱ had statistic differences（F=26.930，P=0.001）. The autophagy level was lower in p-USCs than in USCs（F=15.060，P=0.000）. Conclusion：There were two types of p-USCs in peadiatric patients with simple primary nephrotic syndrome，namely p-USCs Ⅰ and p-USCs Ⅱ. Compared with USCs，the p-USCs has different mor－phological features，lower proliferation，Pax2 expression and au－tophagy level.