摘要
目的原代培养和鉴定大鼠皮层星形胶质细胞,并建立大鼠皮层星形胶质细胞氧糖剥夺(OGD)模型。方法大鼠星形胶质细胞原代培养后,作胶质纤维酸性蛋白(GFAP)免疫细胞化学染色鉴定,应用缺氧D-Hank液及缺氧培养罐行OGD后用台盼蓝染色及乳酸脱氢酶(LDH)漏出率检测细胞损伤程度。结果星形胶质细胞培养至第9天,GFAP染色阳性细胞为98.54%±2.01%;星形胶质细胞经不同时间OGD处理后,台盼蓝染色呈现不同形态;OGD60min时,星形胶质细胞LDH漏出率较对照组明显增加(P<0.05),并随时间递增。结论成功完成大鼠皮层星形胶质细胞原代培养,星形胶质细胞在培养第9天可用于模型制备,成功建立星形胶质细胞OGD模型。
Objective To conduct primary culture and identification of rat cerebral cortical astrocytes,and to set up the oxygen/glucose deprivation （OGD） model. Methods Cells presented in the culture were shown to be astrocytes after characterization by immunocytochemistry,using specific anti-GFAP antibody. OGD model was established with hypoxic D-Hank′s and anaerobic chamber and identified by cell death and LDH release rate. Results Immunohistochemistry of cultured cell showed that the positive rates of GFAP were 98.54%±2.01% at the 9th day;different shape and motility identified by trypan-blue stain was observed when exposed to different OGD time;60 min OGD caused a significant increase on the levels of LDH release （P〈0.05）. Conclusions The primary culture astrocytes could be explored to OGD at the 9th day. The model of OGD ischemia in astrocytes is successfully established by hypoxic D-Hank′s and an anaerobic chamber.
出处
《中国老年学杂志》
CAS
CSCD
北大核心
2010年第3期340-342,共3页
Chinese Journal of Gerontology
基金
教育部高等学校博士学科专项基金项目(20060183053)
吉林省科技厅自然科学基金项目(200705186)
