昆明小鼠胰腺星状细胞分离培养与鉴定  

Isolation,culture and identification of pancreatic stellate cells in Kunming mice

作　　者：许小凡[1] 吴楠[2] 朱林佳[2] 张红[1] 

机构地区：[1]陕西中医药大学医学科研实验中心,陕西咸阳712046 [2]陕西中医药大学基础医学院,陕西咸阳712046

出　　处：《新乡医学院学报》2016年第7期564-567,共4页Journal of Xinxiang Medical University

基　　金：国家自然科学基金资助项目(编号:81102725);陕西省教育厅自然科学重点科研计划项目(编号:15JS027)

摘　　要：目的通过体外分离培养昆明小鼠胰腺星状细胞(PSC),建立更为经济合适的分离PSC方法。方法采用酶消化联合碘海醇密度梯度离心分离小鼠PSC,经油红O染色和荧光倒置显微镜观察鉴定原代培养的PSC,传代并观察其活化状态。结果昆明小鼠分离PSC数量可达1.84×105,存活率为93.0%。经自发荧光及油红O染色鉴定原代分离PSC脂滴,发现PSC纯度较高。形态观察经传代培养的PSC在前3代细胞状态良好,第4代PSC细胞形态呈现活化且α-平滑肌肌动蛋白染色呈阳性。结论酶消化联合碘海醇密度梯度离心可以成功分离昆明小鼠PSC,前3代PSC的活力与纯度均能满足体外实验的要求,昆明小鼠可作为更为经济适用的PSC分离动物。Objective To establish more economical and appropriate model to isolate pancreatic stellate cells in vitro by isolating and culturing pancreatic stellate cells in Kunming mice. Methods The pancreatic stellate cells of mice were isolated by enzymatic digestion and iohexol density gradient centrifugation,the primary cultured pancreatic stellate cells were stained by oil red O and identified by fluorescence microscope. The activated state was observed in the passage cells. Results The number of pancreatic stellate cells isolated from Kunming mice could be 1. 84 × 105,the viability was 93. 0%. The result of autofluorescence and oil red O staining showed that primary pancreatic stellate cells had high purity. Morphological observation showed that the pancreatic stellate cells of the first three passages were in good condition,the generation was activated,and the expression of alpha-sooth muscle actin was positive. Conclusions The pancreatic stellate cells can be isolated successfully by enzymatic digestion and iohexol density gradient centrifugation,and the first three passengers can meet the requirements of experiment in vitro. So Kunming mice can be a more economical and applicable animals for pancreatic stellate cells isolation.

关 键 词：胰腺星状细胞 传代 油红O Α-平滑肌肌动蛋白 昆明小鼠 

分 类 号：Q481[生物学—生理学] R322.491[医药卫生—人体解剖和组织胚胎学]