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用胶原酶消化、差异离心和尼龙网过滤的方法分离鼠脑微血管内皮细胞 ,建立其体外长期培养方法。经形态学 ,免疫组化 ,酶学等鉴定 ,培养细胞为脑微血管内皮细胞。动态观察培养细胞酶含量变化 ,发现随着细胞培养时间的延长 ,血管紧张素转换酶 (ACE)呈上升趋势 ,而γ-谷氨酰胺转化酶 (γ- GT)和硷性磷酸酶 (AL P)则明显下降。实验结果提示 ,血脑屏障的主要功能酶γ- GT和 AL P可作为体外脑微血管内皮细胞的标志酶 ,但要维持长时间体外表达则需要某种因子的介导。本实验可为体外研究血脑屏障及相关疾病提供帮助。
The rat brain microvascular endothelial cells were isolated by collagenase digestion, differential centrifugation and nylon mesh filtration to establish their long-term culture method in vitro. The morphological, immunohistochemical, and enzymatic studies identified the cultured cells as brain microvascular endothelial cells. Observing the change of enzyme content in cultured cells, we found that with the prolongation of cell culture time, angiotensin-converting enzyme (ACE) showed an upward trend, while γ-glutamine converting enzyme (γ-GT) and alkaline phosphatase (ALP). ) It dropped significantly. The experimental results suggest that the main functional enzymes of the blood-brain barrier, γ-GT and AL P, may serve as marker enzymes for in vitro brain microvascular endothelial cells, but it requires mediation of certain factors to maintain prolonged in vitro expression. This experiment can help to study blood-brain barrier and related diseases in vitro.