皮肤血管瘤内皮细胞原代分离培养与鉴定  

Skin Hemangioma Endothelial Cells Original Generation of Isolated Culture and Appraisal

作　　者：饶国洲[1] 李昂[1] 张引成[2] 孙惠玲[1] 朱永进[1] 

机构地区：[1]西安交通大学口腔医院中心实验室,西安710004 [2]西安交通大学口腔医院口腔颌面外科,西安710004

出　　处：《现代检验医学杂志》2009年第5期64-66,共3页Journal of Modern Laboratory Medicine

摘　　要：目的建立一种体外分离纯化血管瘤内皮细胞原代培养方法，探讨血管瘤内皮细胞的生物学特性及其分子机制。方法采用组织块法结合差速贴壁进行原代培养。通过相差显微镜观察细胞形态，免疫组织化学和免疫荧光染色鉴定细胞CD34和Ⅶ因子相关抗原。结果培养48h后可见组织块周围游离出细胞，呈多边形，胞核清晰，72h去除组织块，2w后细胞长满培养板底形成单层细胞呈铺路石样生长。免疫组化显示CD34表达阳性；免疫荧光染色证实Ⅶ因子相关抗原阳性。结论组织块法结合差速贴壁可获得高纯度的血管瘤内皮细胞，是一种简单实用的原代培养方法。Objective To establish a cell culture method which is isolation and purification of hemangioma endothelial cells and discuss the biological characteristics and molecular mechanism of hemangioma endothelial cells. Methods Primary culture combined primary tissue culture and differential adhesion. Through the phase contrast microscope to observe cell morphology,identify CD34 cell and Ⅶ factor-related antigen expression by the way of immunohistochemistry and immunofluorescence. Results After 48h culture could be seen free cells which were surrounding tissue. The cells were polygonal and had clear nucleus. Removal of tissue after 72h. After 2 weeks the cell plate was covered with cells at the end of the formation of single-layer paving stone-like growth. Immunohistochemistry showed expression of CD34 was positive. Immunofluorescent staining confirmed Ⅶ factor-related antigen was positive. Conclusion Combination of primary tissue culture and differential adhesion can obtain high purity hemangioma endothelial cells. It is a simple and practical method of primary culture.

关 键 词：血管瘤 细胞培养 免疫组化 细胞鉴定 

分 类 号：R732.2[医药卫生—肿瘤] Q813.1[医药卫生—临床医学]