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取大鼠周边肺组织进行肺微血管内皮细胞培养。将肺组织切成小块,用含有20%新生牛血清、肝素90μg/ml、L-谷胺酰胺4mmol、青霉素100U/ml和链霉素100μg/ml的RPMI-1640培养基培养。血细胞立即从肺组织周围游出。继而是肺微血管内皮细胞。成纤维细胞及其他细胞72h才游出。培养60h后取出肺组织块,培养瓶中只有微血管内皮细胞和血细胞。后者可通过传代除去。获得的肺微血管内皮细胞具有规律的鹅卵石样形态和对异植物血凝素结合试验及八因子相关抗原免疫荧光染色均阳性。
Peripheral lung tissue was taken for pulmonary microvascular endothelial cell culture. The lung tissue was cut into small pieces and cultured in RPMI-1640 medium containing 20% fetal bovine serum, heparin 90 μg / ml, L-glutamine 4 mmol, penicillin 100 U / ml and streptomycin 100 μg / ml. Blood cells immediately swim around the lung tissue. Followed by pulmonary microvascular endothelial cells. Fibroblasts and other cells 72h before swimming out. After 60 hours of culture, the lung tissue was removed and only the microvascular endothelial cells and blood cells were cultured in the flask. The latter can be removed by passage. The obtained pulmonary microvascular endothelial cells have a regular pebble-like morphology and are positive for allo-hemagglutinin binding assay and eight-factor-related antigen immunofluorescence staining.