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从糖尿病自发性突变(Lepr/db)纯合子小鼠肝脏分离出肝窦内皮细胞,表现为病理性肥胖、慢性高血糖、胰腺细胞萎缩和降血脂症。BKS db小鼠肝窦内皮细胞培养于T25组织培养瓶中,预先包覆以明胶为基础的溶液,培养2 min,在AcceGen’s culture Complete Growth Medium中培养,一般扩增培养3-7天。在运输之前,第3代的细胞从培养瓶中分离出来,立即冷冻保存在小瓶中。每瓶每毫升至少含有0.5×106个细胞。我们分离原代内皮细胞的方法是在已有方法和我们的专利方法的基础上发展起来的。这些细胞被预涂PECAM-1抗体筛选,随后应用预涂二抗的磁珠分选。
BKS db Mouse Liver Sinusoidal Endothelial Cells are isolated from the liver of Mice homozygous for the diabetes spontaneous mutation (Lepr/db) manifest morbid obesity, chronic hyperglycemia, pancreatic beta cell atrophy and become hypoRIemic. BKS db Mouse Liver Sinusoidal Endothelial Cells are grown in T25 tissue culture flasks pre-coated with gelatin-based solution for 2 min and incubated in AcceGen's Culture Complete Growth Medium generally for 3-7 days. Cultures are then expanded. Prior to shipping, cells at passage 3 are detached from flasks and immediately cryopreserved in vials. Each vial contains at least 0.5×106 cells per ml. The method we use to isolate primary endothelial cells was developed based on a combination of established and our proprietary methods. These cells are pre-coated with PECAM-1 antibody, following the application of magnetic beads pre-coated with secondary antibody
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