小鼠肝脏中细胞角蛋白19阳性细胞的肝向分化特性

发布时间：2018-03-03 17:19

本文选题：肝干细胞　切入点：CK　出处：《第二军医大学学报》2017年05期 　论文类型：期刊论文

【摘要】：目的明确小鼠肝脏中的细胞角蛋白19(cytokeratin 19,CK19)阳性(CK19~+)细胞是否可分化为成熟肝细胞。方法利用CK19~(CreERT)小鼠和Rosa26-GFP小鼠杂交得到CK19~(CreERT)/Rosa26-GFP双转基因小鼠,注射他莫昔芬(tamoxifen,TM)后检测小鼠肝脏中CK19~+细胞的GFP标记情况。在此基础上分别构建3,5-二乙氧基羰基-1,4-二氢-2,4,6-三甲基吡啶和四氯化碳(carbon tetrachloride,CCl4)肝脏损伤模型,采用肝脏组织冰冻切片结合免疫荧光染色检测肝脏中GFP标记的CK19~+细胞的分化情况。结果获得CK19~(CreERT)/Rosa26-GFP双转基因小鼠,免疫荧光染色结果显示CK19~+细胞可被GFP标记。在DDC肝脏损伤模型小鼠中检测到增生性小胆管内有GFP~+细胞,这些GFP~+细胞表达胆管上皮细胞标志物CK19,且DDC肝损伤模型小鼠中GFP~+胆管细胞比例高于未损伤对照组[(63.5±6.3)%vs(53.6±4.8)%,P0.05];在CCl4肝损伤模型组小鼠中检测到肝脏实质细胞中有GFP~+细胞,这些GFP~+细胞表达成熟肝细胞标志物白蛋白(ALB),且CCl4肝损伤模型组小鼠肝脏中GFP~+细胞比例高于未损伤对照组[(0.15±0.02)%vs(0.008±0.003)%,P0.01]。结论小鼠肝脏内的CK19~+细胞群体中存在具有肝向分化潜能的前体细胞,可能为真正的肝干细胞的识别鉴定提供一个新线索。
[Abstract]:Objective to determine whether cytokeratin 19 cytokeratin 19 (CK19) positive cells can differentiate into mature hepatocytes in mouse liver. Methods CK19~(CreERT)/Rosa26-GFP double transgenic mice were obtained by hybridization with Rosa26-GFP mice. The GFP labeling of CK19 ~ ~ cells in mouse liver was detected after tamoxifende (TMN) injection. On the basis of this, the liver injury models of 3O5-diethoxycarbonyl-4-dihydro-4-dihydro-6-trimethylpyridine and carbon tetrachloridede CCl4 were constructed, respectively. The differentiation of CK19 ~ ~ cells labeled with GFP in liver was detected by using frozen sections of liver tissue and immunofluorescence staining. Immunofluorescence staining showed that CK19 ~ ~ cells could be labeled by GFP. GFP~ cells were found in proliferative small bile duct in DDC liver injury model mice. These GFP~ cells expressed CK19, and the proportion of GFP~ cells in DDC liver injury model mice was higher than that in the uninjured control group [63.5 卤6.3% vs 53.6 卤4.8]. GFP~ cells were detected in liver parenchymal cells in CCl4 liver injury model group. The ratio of GFP~ cells in CCl4 liver injury model group was higher than that in uninjured control group [0.15 卤0.02 卤0.008 卤0.003] .Conclusion there are liver differentiation potential progenitor cells in CK19~ ~ cell population of mouse liver, and the expression of ALB ~ + in GFP~ + cells is higher than that in normal control group [0. 15 卤0. 02% vs 0. 008 卤0. 003]. It may provide a new clue for the identification of true liver stem cells.
【作者单位】：第二军医大学基础部细胞生物学教研室;第二军医大学国际合作肿瘤研究所;
【基金】：国家自然科学基金(31401166,81670573) 国家重点基础研究发展计划(“973计划”,2010CB945602)~~
【分类号】：R575

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