You Position: Home > Paper

Fresh isolation and identification of the rat liver Kupffer cells suitable to patch clamp technique

( views:0, downloads:0 )
Author:
No author available
Journal Title:
WORLD CHINESE JOURNAL OF DIGESTOLOGY
Issue:
2
DOI:
10.3969/j.issn.1009-3079.2005.02.012
Key Word:
Kupffer细胞;膜片钳技术

Abstract´╝Ü 目的:建立一种适于膜片钳技术(PCT)的大鼠肝Kupffer细胞的急性分离方法.方法:经门静脉插管,用无Ca2+,Mg2+K-H原位灌注液冲洗后,Ⅳ型胶原酶原位循环灌注消化,Percoll不连续密度梯度离心,分离制备大鼠肝Kupffer细胞,选择性贴壁纯化.用体内、体外吞噬功能(炭素墨水或聚苯乙烯乳珠)实验鉴定制备的肝Kupffer细胞.用全细胞膜片钳记录技术测定Kupffer细胞电流.结果:分离获得的大鼠肝Kupffer细胞形态多样,典型的为多角形或星形,细胞分离产量为2.5-3.5×106/g,细胞纯度90%,贴壁率为39.4%.用台盼蓝染色鉴定,细胞活性大于90%,多数细胞明显吞噬颗粒.易用于PCT,且记录到钙池操纵的Ca2+通道电流(store-operated Ca2+channel currents,Isoc).结论:用胶原酶原位循环灌注消化,结合Percoll不连续密度梯度离心和选择性贴壁纯化,成功分离制备出高产、纯度高的适于PCT的肝Kupffer细胞.

WanfangData CO.,Ltd All Rights Reserved
About WanfangData | Contact US
Healthcare Department, Fuxing Road NO.15, Haidian District Beijing, 100038 P.R.China
Tel:+86-010-58882616 Fax:+86-010-58882615 Email:yiyao@wanfangdata.com.cn