大鼠肾乳头组织干细胞的分离与鉴定**

背景：迄今为止尚未见到从活体肾乳头部位提取肾组织干细胞及对其细胞性状特点进行分析的研究。 目的：在体外分离培养和鉴定大鼠肾乳头部组织干细胞,并将其生物学特性与骨髓间充质干细胞进行比较。 方法：从SD雄性大鼠肾脏分离提取肾乳头尖部细胞,从股骨胫骨的骨髓腔提取骨髓细胞分别接种于培养瓶,加入含体积分数为10%胎牛血清的DMEM/F12培养基进行体外培养并传代,取对数生长期的第3代细胞行成脂、成骨诱导分化。 结果与结论：①大鼠肾乳头部组织干细胞呈玫瑰花形旋涡状生长,多呈短梭形和多边形。②细胞的生长曲线呈“S”型。③细胞经成脂诱导后油红O染色阳性;成骨诱导茜素红染色后可见橘红色钙盐沉积。④流式细胞术检测显示肾乳头部组织干细胞表达CD29,CD44,CD90阳性,CD45阴性。结果证实肾乳头部位存在具有间充质干细胞特性的组织干细胞。关键词：肾脏;乳头部;组织干细胞;诱导分化;间充质干细胞 doi:10.3969/j.issn.1673-8225.2012.19.017

Isolation and identification of rat renal papilla stem cells

BACKGROUND: There is little research about isolation of renal stem cells from the renal papillary in living donor and the characteristics of the cells. OBJECTIVE: To isolate and identify the renal papilla stem cells in vitro, and compare their biological characteristics to bone marrow mesenchymal stem cells. METHODS: Cells in the tip of the renal papilla in Sprague-Dawley rats were isolated, and bone marrow mesenchymal stem cells were from the bone marrow cavities of femur and tibia. The cells were seeded in culture flasks respectively. DMEM/F12 media containing 10% fetal bovine serum were added into the culture flasks for culture and passage in vitro. When 70%-80% confluence, 0.25% Trypsin were used to digest and passage the cells. The third-passage cells were used for adipogenic and osteogenic differentiation. RESULTS AND CONCLUSION: The renal papilla stem cells showed a rose-shaped and vortex-like growth and the cells were short spindle and polygons shaped. The growth curves of the cells were “S”-shape. The cells showed positive staining by oil red O after adipogenic differentiation, and orange calcium deposition could be seen in the cells by alizarin red staining after osteogenic differentiation. The flow cytometer analysis showed that CD29, CD44 and CD90 positively expressed in these cells, while CD45 negatively expressed in them. It is indicated that tissue stem cells containing the characteristics of mesenchymal stem cells exist in the renal papilla.