【作者】 张贝莹； 罗冬章； 麦海涛； 王丙云； 陈胜锋； 陈志胜； 计慧琴； 冼琼珍；

【Author】 Zhang Beiying;Luo Dongzhang;Mai Haitao;Wang Bingyun;Chen Shengfeng;Chen Zhisheng;Ji Huiqing;Xian Qiongzhen;Department of animal medicine,Foshan University;

【机构】 佛山科学技术学院生命科学与工程学院；

【摘要】 本实验采用密度梯度离心法对犬骨髓液进行分离,获取单个核细胞。通过培养纯化获得骨髓间充质干细胞,并分别对第3和第6代的犬骨髓间充质干细胞进行成脂诱导分化。结果表明,犬骨髓间充质干细胞原代培养呈椭圆形或多角形,第4d细胞贴壁,第6d形成细胞集落,第12d细胞呈长梭形、排列有明显方向性,细胞排列成漩涡状、网状或辐射状。传代细胞倍增速度比原代细胞明显增快。细胞传至7代,其形态保持不变。油红0染色后可见诱导组细胞被染红的脂滴,而对照组没被染色,提示犬骨髓间充质干细胞被诱导分化为脂肪细胞;比较第3和第6代的犬骨髓间充质干细胞诱导分化结果,第3代的犬骨髓间充质干细胞形成的脂滴比第6代的形成的脂滴多,提示间充质干细胞诱导分化的能力随传代次数的增多而减弱。更多还原

【Abstract】 To explore a convenient,strong operability method to separate and culture a high purity of bone marrow mesenchymal stem cells from dogs,then identify the cells,in order to provide experimental basis for applications in the future.This experiment adopted the density gradient centrifugation to obtain mononuclear cells.Ficoll was used to separate mononuclear cells from bone marrow fluid,BMSCs was obtained by culture purification,then identifying the cells of P3 and P6 by in vitro adipogenic differentiation and oil red O staining.The results showed that Dog BMSCs of primary culture presented ellipse or polygonal shape.Most cells adhered at day 4,the cell colonies were visible at day 6,cells showed long spindle and had obvious directionality in circinate,reticular and radial shape.Dog BMSCs of continuous culture grew faster than primary culture.Dog BMSCs’ shape remained the same after culturing to 7 generation.Red lipid drops could be visible in experimental group,but the control couldn’t,it meant dog BMSC can be induced to differentiate into adipocyte.Comparing the adipogenic differentiation of dog DMSCs between P3 and P6,the visible lipid drops of P3 were more than P6.It showed that BMSCs differentiation ability weakened with the number of batches increasing.更多还原