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目的 :培养的内皮细胞为基础和临床研究提供体外模型 ,并为进一步研究内皮细胞对造血的影响提供实验材料。方法 :经胶原酶消化获得人脐静脉内皮细胞 ,用RPMI 164 0加 2 0 %的胎牛血清进行培养。结果 :成簇状的内皮细胞最初形成于培养皿 (或培养瓶 )的底层 ,继而细胞迅速生长 ,融合成片 ,约 7天细胞呈单层多角形镶嵌排列。透射电镜观察 ,原代培养细胞的胞质内含内皮细胞特有的细胞器 (WPBs)。兔抗人Ⅷ因子相关抗原 (ⅧR∶Ag)免疫组织化学染色实验证实 ,培养的内皮细胞含有ⅧR∶Ag。 结论 :本实验的各项结果从形态 ,免疫组化及透射电镜的观察均符合内皮细胞的特征 ,因此确信所培养的内皮细胞可提供作进一步深入的细胞生物学及对造血影响的研究。
OBJECTIVE: Cultured endothelial cells provide in vitro models for both basic and clinical studies and provide experimental materials for further study of the effects of endothelial cells on hematopoiesis. Methods: Human umbilical vein endothelial cells were obtained by collagenase digestion and cultured with RPMI160 + 20% fetal bovine serum. Results: Clustered endothelial cells were initially formed on the bottom of petri dishes (or culture flasks), followed by rapid cell growth and fusion to form pieces with approximately monolayer polygon inlaid arrangement at about 7 days. Transmission electron microscopy showed that the cytoplasm of primary cultured cells contained endothelial cell-specific organelles (WPBs). Rabbit anti-human Ⅷ factor-related antigen (Ⅷ R: Ag) immunohistochemical staining experiments confirmed that cultured endothelial cells contain Ⅷ R: Ag. CONCLUSIONS: The results of this experiment are morphologically, immunohistochemically, and TEM observations consistent with the characteristics of endothelial cells and it is therefore believed that cultured endothelial cells may provide further insights into cell biology and the effects on hematopoiesis.