[1]许小凡,吴　楠,朱林佳,等.昆明小鼠胰腺星状细胞分离培养与鉴定[J].新乡医学院学报,2016,33(7):564-567.[doi:10.7683/xxyxyxb.2016.07.004]
　XU Xiao-fan,WU Nan,ZHU Lin-jia,et al.Isolation，culture and identification of pancreatic stellate cells in Kunming mice[J].Journal of Xinxiang Medical University,2016,33(7):564-567.[doi:10.7683/xxyxyxb.2016.07.004]
点击复制

昆明小鼠胰腺星状细胞分离培养与鉴定
分享到：

《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
33
期数:
2016年7
页码:
564-567
栏目:
基础研究
出版日期:
2016-07-05

文章信息/Info

Title:
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
（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）
Author(s):
XU Xiao-fan1; WU Nan2; ZHU Lin-jia2; ZHANG Hong1
(1.Medical Research Center，Shaanxi University of Chinese Medicine，Xianyang 712046，Shaanxi Province，China；2.College of Basic Medicine，Shaanxi University of Chinese Medicine，Xianyang 712046，Shaanxi Province，China)
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
Keywords:
pancreatic stellate cell; passage; oil red O; alpha-sooth muscle actin; Kunming mice
分类号:
Q481，R322.4+91
DOI:
10.7683/xxyxyxb.2016.07.004
文献标志码:
A
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。
Abstract:
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.Conclusion　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.

参考文献/References:

［1］　APTE M，PIROLA R C，WILSON J S.Pancreatic stellate cell：physiologic role，role in fibrosis and cancer［J］.Curr Opin Gastroenterol，2015，31(5)：416-423.
［2］　MASAMUNE A，WATANABE T，KIKUTA K，et al.Roles of pancreatic stellate cells in pancreatic inflammation and fibrosis［J］.Clin Gastroenterol Hepatol，2009，7(Suppl 11)：S48-S54.
［3］　张晓芹，刘芳，张红.慢性胰腺炎发病机制研究进展［J］.新乡医学院学报，2014，31(2)：142-145.
［4］　崔立华，张淑坤，崔乃强，等.大鼠胰腺星状细胞分离培养与鉴定［J］.中国中西医结合外科杂志，2014，20(5)：504-507.
［5］　PHILLIPS P A，MCCARROLL J A，PARK S，et al.Rat pancreatic stellate cells secrete matrix metalloproteinases：implications for extracellular matrix turnover［J］.Gut，2003，52(2)：275-282.
［6］　唐元瑜，水楠楠.油红O染色原代未活化胰腺星状细胞脂肪滴的实验观察［J］.中国组织化学与细胞化学杂志，2013，22(3)：266-268.
［7］　MASAMUNE A，SHIMOSEGAWA T.Pancreatic stellate cells：multi-functional cells in the pancreas［J］.Pancreatology，2013，13(2)：102-105.
［8］　APTE M，PIROLA R，WILSON J.The fibrosis of chronic pancreatitis：new insights into the role of pancreatic stellate cells［J］.Antioxid Redox Signal，2011，15(10)：2711-2722.
［9］　APTE M V，HABER P S，APPLEGATE T L，et al.Periacinar stellate shaped cells in rat pancreas：identification，isolation，and culture［J］.Gut，1998，43(1)：128-133.
［10］　范昕，王莹，侯雯跻，等.小鼠胰腺星状细胞的分离培养与鉴定［J］.江苏大学学报（医学版），2013，22(6)：468-471.
［11］　唐元瑜，苏式兵.小鼠胰星状细胞的分离培养及鉴定［J］.世界华人消化杂志，2010，18(1)：28-32.

相似文献/References:

[1]姬明丽１,王煜霞２.一种挽救轻度污染的肿瘤贴壁细胞的方法[J].新乡医学院学报,2015,32(09):814.

更新日期/Last Update: 2016-07-05