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目的 探讨脑微血管内皮细胞的体外培养方法并进行细胞超微结构研究及组织型纤溶酶原激活物(TPA)活性测定。 方法 取新生小鼠脑组织 ,通过匀浆、过筛、胶原酶消化、差速粘附等技术对鼠脑微血管内皮细胞进行原代培养 ,待细胞铺满瓶底时 ,用 0 .12 5 %胰酶 - 0 .0 2 % EDTA消化 ,离心收集内皮细胞 ,进行传代培养。原代、传代各取 8例 ,吸取培养液用酶联免疫吸附试验测试 TPA活性。 结果 经 因子相关抗原免疫组织化学鉴定、细胞超微结构观察 ,证明培养的是血管内皮细胞。培养的脑微血管内皮细胞能合成分泌 TPA。 结论 鼠脑微血管内皮细胞的培养为体外研究脑血管疾病提供了可靠的手段。
Objective To investigate the in vitro culture method of brain microvascular endothelial cells and to study the ultrastructure and the activity of tissue plasminogen activator (TPA). Methods The brain tissue of newborn mice was taken for primary culture of mouse brain microvascular endothelial cells by homogenization, sieving, collagenase digestion and differential adhesion. When the cells were covered with bottom, Trypsin - 0. 2% EDTA digestion, endothelial cells were collected by centrifugation, subculture. The primary passage, the passage of each take 8 cases, the culture medium was tested by enzyme-linked immunosorbent assay TPA activity. Results Immunohistochemical identification of factor-related antigens, cell ultrastructure observation, to prove that the culture of vascular endothelial cells. Cultured brain microvascular endothelial cells can synthesize and secrete TPA. Conclusion The culture of rat brain microvascular endothelial cells provides a reliable means of studying cerebrovascular diseases in vitro.