期刊论文

Primary cultivation and purification of rat cortical astrocytes and cell identification by detecting of GFAP by HCA

中文

沈阳药科大学学报

1006-2858

2016

33

7

581-587

10.14066/j.cnki.cn21-1349/r.2016.07.013

81573965:国家自然科学基金 81403379:国家自然科学基金 ZYFT201401:湖南省中药粉体与创新药物省部共建国家重点实验培育基地开放基金资助项目

本校为第一机构

目的探索大鼠脑皮质星形胶质细胞(astrocyte,AS)的分离、原代培养及纯化方法,采用高内涵细胞成像分析技术(high content analysis,HCA)对其进行胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)鉴定。方法取2、3 d的新生SD大鼠,体视显微镜下获取大脑皮质,机械分离成1 mm3组织碎块,采用质量分数为0. 25%的胰蛋白酶和0. 2%的胶原酶(体积比为1∶ 1)消化后进行体外培养,分别观察接种1 h、3 d、5 d后原代AS基本形态结构,酶消化3 min后,接种30 min,5 d后传代AS基本形态结构; 采用多次、多阶段的差速贴壁和震荡相结合的方法纯化细胞; 采用HCA技术对AS进行GFAP免疫细胞化学鉴定。结果采用本法培...

Objective To establish a method of separation,primary cultivation and purification for rat cortical astrocytes and to identify the cell by detecting GFAP with high content analysis. Methods Astrocytes were obtained from 2 - 3 days old neonatal Sprague-Dawley rat under stereoscopic microscope,and digested by trypsin(0. 25%) -collagenase(0. 2%) (V∶ V = 1∶ 1) after been broken mechanically into tissue fragments with volume of Ca. 1mm3. And then cells were seeded in cell culture flask to observe its basic morphology structure at 1h,3 day,5 day af...