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目的探索犬肢体血管内皮细胞简易培养方法，为在肢端复合组织体 外保存方法及保存后组织细胞（血管内皮细胞）活性判定方面的研究奠定基础。方 法手术切取犬旋股下动脉侧支，采用血管外翻术，消化分离血管内皮细胞后体 外培养。用免疫组化法对增殖细胞分别进行Ⅷ因子和ＣＯ３４相关抗原的检测。结果 分离的细胞存活率为 ９９．３％。培养 ２４ｈ多数细胞已贴壁，至第３ｄ，细胞开始增殖，可见细胞分裂相；至第７ｄ，亚细胞克隆组单个细胞分裂增殖，呈环片样生长； 扩增组细胞增殖旺盛，是铺路石样结构。Ⅷ因子和ＣＤ３４相关抗原表达阳性。结论 血管内皮细胞的分离采用血管外翻术后消化可获理想的分离效果，该方法简单易 行、重复性好；体外培养的犬血管内皮细胞可以检测到Ⅷ因子和ＣＤ３４相关抗原。
Objective To explore the method of simple culture of vascular endothelial cells in canine limbs and lay a foundation for further study on the method of preserving the composite tissues of extremities in vitro and the determination of the activity of tissue cells (vascular endothelial cells) after preservation. Methods The circumcision of the circumflex femoral artery was excised surgically. The vessel was everted and the endothelial cells were digested and separated for culture in vitro. Immunohistochemistry was used to detect the proliferating cells respectively Ⅷ factor and CO34 related antigen. Results The cell survival rate was 99.3%. After cultured for 24 hours, most of the cells were adherent to the third day. The cells began to proliferate and the cell division phase was seen. On the 7th day, single cell subcellular clonal division proliferated in a ring-like manner. Sample structure. Ⅷ factor and CD34-associated antigen expression was positive. Conclusions The separation of vascular endothelial cells by extravascular angioplasty can achieve satisfactory separation effect. The method is simple and reproducible, and can detect Ⅷ factor and CD34 related antigen in cultured canine vascular endothelial cells.