You Position: Home > Paper
Isolation, culture and identification of mouse pancreatic satellite cells
( views:190, downloads:660 )
- Author:
- No author available
- Journal Title:
- WORLD CHINESE JOURNAL OF DIGESTOLOGY
- Issue:
- 1
- DOI:
- 10.3969/j.issn.1009-3079.2010.01.005
- Key Word:
- 胰星状细胞;培养;鉴定;植块法
Abstract： 目的:建立简单易行的小鼠胰星状细胞(PSCs)原代培养方法,为胰腺纤维化研究提供可靠的体外细胞模型.方法:采用植块法结合酶消化法进行培养,即在无菌条件下取小鼠正常胰腺组织,经过剪碎、胰蛋白酶消化后植入培养瓶中贴壁培养,并予限制性条件培养基进行纯化,通过倒置生物显微镜对传代前后所培养细胞进行形态学、自发荧光及油红染色脂滴的观察,并结合细胞免疫化学和免疫荧光等方法来鉴定小鼠PSCs.结果:小鼠原代PSCs培养3-4 d后油红染色阳性,并能观察到自发荧光现象;传代后的细胞形态主要表现为体积较大,伪足发达,呈"星芒状"或"乌贼样";细胞免疫荧光染色和细胞免疫化学染色显示α-平滑肌肌动蛋白(α-SMA)表达阳性.结论:植块与酶消化结合法分离小鼠胰腺星状细胞,简单实用、成功率高,能够满足体外实验的要求.
- [1]吴恺,徐刚,王兴鹏.甘草酸对胰腺星状细胞细胞外基质代谢调节作用的研究[J].2007,(11).
- [2]贾一韬,李兆申.大鼠胰星状细胞的分离与培养[J].2003,(3).
- [3]周龙安,杨桂元,钱祝银.腹主动脉灌注法分离胰腺星状细胞及其培养、鉴定[J].2007,(1).
- [4]万远太,赵秋,王天才,等.大鼠胰星状细胞体外分离培养的实验研究[J].2005,(1).
- [5]主编李兆申, 许国铭.现代胰腺病学[M].2006.
- [6]袁耀宗.胰腺病学新进展与新技术[M].2000.
- [7]Hildebrand PB,Schmidt WE,Kruse ML,etc.Isolation, long-term culture, and characterization of rat pancreatic fibroblastoid/stellate cells.[J].2001,23(1).
- [8]Max G. Bachem*,Erik Schneider*,Hans Groß*,etc.Identification, culture, and characterization of pancreatic stellate cells in rats and humans[J].1998,115(2).
- [9]Chen Y,Kleeff J,Lohr M,etc.Immortalization of pancreatic stellate cells as an in vitro model of pancreatic fibrosis: deactivation is induced by matrigel and N-acetylcysteine.[J].2005,85(10).
- [10]M V Apte,P S Haber,T L Applegate,etc.Periacinar stellate shaped cells in rat pancreas: identification, isolation, and culture[J].1998,43(1).