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 |
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引用本文: | 魏爱宣,李昕华,闫世军,何金婷,莽靖,邢影,邵延坤,徐忠信. 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[J]. 中国老年学杂志, 2010, 30(3) |
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作者姓名: | 魏爱宣 李昕华 闫世军 何金婷 莽靖 邢影 邵延坤 徐忠信 |
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作者单位: | 1. 吉林市中心医院神经内科 2. 长春市中心医院神经内科 3. 吉林大学第二医院神经外科 4. 吉林大学中日联谊医院神经内科,吉林,长春,130033 |
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基金项目: | 教育部高等学校博士学科专项基金项目,吉林省科技厅自然科学基金项目 |
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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;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.
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关 键 词: | 星形胶质细胞 原代培养 氧糖剥夺(OGD) |
Establishment and identification of oxygen glucose deprivation model of primary culture rat cortical astrocytes |
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Abstract: | 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. |
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Keywords: | Astrocytes Primary culture Oxygen glucose deprivation (OGD) |
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