【作者】 冼伟杭； 何诗； 王丙云； 李东升； 阮慧敏； 叶彩玲； 李星颖； 陈胜锋； 白银山； 陈志胜； 刘璨颖； 计慧琴；

【Author】 XIAN Weihang;HE Shi;WANG Bingyun;LI Dongsheng;RUAN Huiming;YE Cailing;LI Xingying;CHEN Shengfeng;BAI Yinshan;CHEN Zhisheng;LIU Canying;JI Huiqin;collage of Life Science and Engineering,Foshan University;Vetcell biotechnology limited company;

【机构】 佛山科学技术学院生命科学与工程学院； 维赛(佛山)生物科技有限公司；

【摘要】 肾急性损伤是犬常发疾病,目前常规治疗无法完全恢复肾功能,影响犬的健康。本实验通过Ⅰ型胶原酶消化法分离获得犬肾干细胞并进行传代培养;采用CCK8法测定细胞活性并绘制生长曲线;利用流式细胞术对细胞表面标记物进行鉴定;利用PCR法扩增CD133特异表面标记物基因;通过成骨、成脂肪诱导分化实验测定其分化能力。实验结果发现:犬肾干细胞呈短梭状,生长曲线呈"S"型;流式细胞术结果为阳性表达CD90和CD29,不表达CD34;PCR成功扩增CD133特异性表面标记物基因。诱导分化实验表明,犬肾干细胞具有成骨、成软骨能力。综上结果表明,本实验成功分离获得犬肾干细胞。更多还原

【Abstract】 Acute kidney injury is a common disease in dogs.At present,conventional treatment can not completely restore renal function,which affects the health of dogs.In this experiment,canine kidney stem cells were isolated and cultured by collagenase digestion method.The cell activity was measured by CCK8 method and the growth curve was plotted.The cell surface markers were identified by flow cytometry.CD133 specific surface marker gene was amplified by PCR.The differentiation ability was determined by osteogenesis and adipogenesis induction differentiation experiments.The results showed that canine renal stem cells were spindle-shaped and the growth curve was S-shaped.Flow cytometry showed positive expression of CD90 and CD29,but no expression of CD34.CD 133 specific surface marker gene was successfully amplified by PCR.The induced differentiation experiment showed that canine renal stem cells had osteogenic and chondrogenic abilities.The results indicated that canine kidney stem cells were successfully isolated in this experiment.Renal stem cells have the function of repairing damaged kidney tissue and restoring renal function,which is expected to be a new therapy for canine kidney injury.更多还原