CAJ | 학술논문

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.
목적 원대배양화감정대서피층성형효질세포,병건립대서피층성형효질세포양당박탈(OGD)모형.방법 대서성형효질세포원대배양후,작효질섬유산성단백(GFAP)면역세포화학염색감정,응용결양D-Hank액급결양배양관행OGD후용태반람염색급유산탈경매(LDH)루출솔검측세포손상정도.결과 성형효질세포배양지제9천, GFAP염색양성세포위98.54%±2.01%;성형효질세포경불동시간OGD처리후,태반람염색정현불동형태;OGD60 min시,성형효질세포LDH루출솔교대조조명현증가(P<0.05),병수시간체증.결론 성공완성대서피층성형효질세포원대배양,성형효질세포재배양제9천가용우모형제비,성공건립성형효질세포OGD모형.
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.