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目的 探讨中枢神经系统海绵状血管瘤(CMs)内皮细胞的分离、培养及鉴定方法。方法 采用手术获得人脑CMs组织35例,通过显微分离、胶原酶消化等程序分离其内皮细胞,相差显微镜下观察其生长状态,CD_(34)、抗平滑肌抗体(α-SMA)免疫荧光染色分别行性状鉴定及纯度检测,透射电镜观察超微结构 结果 10例(67％)未受术中双极电凝影响的标本培养成功,培养 3周后形成内皮细胞单层,细胞形态可分为 2种;平均每隔 7～10 d传代 1次,最多传 4～5代。传代细胞CD_(34)染色呈均一阳性,α-SMA呈阴性。透射电镜下,CMs内皮细胞核呈幼稚状态。结论 获得纯度较高CMs内皮细胞的新方法已经成功建立。
Objective To investigate the methods of isolation, culture and identification of endothelial cells of central nervous system cavernous hemangiomas (CMs). Methods Thirty-five human CMs were obtained by surgery. Endothelial cells were isolated by micro-isolation and collagenase digestion. The growth of the cells was observed under phase contrast microscope. CD34 and α-SMA immunofluorescence staining Characterization and purity test were performed respectively. Ten cases (67%) of the ultrastructural results were observed by transmission electron microscopy. The specimens without bipolar coagulation were successfully cultured. After 3 weeks of culture, endothelial cells were formed. The morphology of the cells could be divided into 2 species; the average every 7 ~ 10 d passage 1, up to pass 4 to 5 generations. Passaged cells CD_ (34) staining showed a positive, α-SMA was negative. Under transmission electron microscope, the nuclei of endothelial cells of CMs were naive. Conclusion A new method to obtain high purity CMs endothelial cells has been successfully established.