大鼠皮层星形胶质细胞原代培养、鉴定及氧糖剥夺模型的建立

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;OGD60 min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.

Establishment and identification of oxygen glucose deprivation model of primary culture rat cortical astrocytes

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.