Curti T, Pasquinelli G, Preda P, Freyrie A, Laschi R, D'Addato M
Department of Vascular Surgery, University of Bologna, Italy.
Ann Vasc Surg. 1989 Oct;3(4):351-63. doi: 10.1016/S0890-5096(06)60159-3.
Human adult endothelial cells were enzymatically harvested from adipose tissue. Cell viability was established by Trypan blue exclusion and transmission and scanning electron microscopy. Endothelial cells were identified by immunocytochemical investigation at light microscopy, transmission electron microscopy, and scanning electron microscopy. Isolated cells were positive for actin and vimentin, negative for desmin. Factor VIII RA was mainly expressed at cell surface and occasionally disclosed in the cytoplasm. Reactivity for UEA I and J15 was weak or undetectable. Human endothelial cells were seeded and left to adhere for one hour onto different nonvascular substrates (glass, poly-l-lysine, formvar-carbon, fibronectin, Teflon). Scanning electron microscopy defined surface features, suggesting tenacious cell adhesion on the substrate. Different vascular substrates were tested (preclotted Dacron, albumin Dacron, Hemashield Dacron, Gelseal Dacron, ePTFE, fibronectin-ePTFE). Commercially available coated grafts showed qualitative and quantitative differences in cell adhesion. In particular, Gelseal Dacron provided the best quantitative results, even though a wide variability was observed. In contrast, fibronectin-coated ePTFE gave more reliable results and high spreading efficiency. In the short term, coated grafts do not seem to offer greater advantages than fibronectin-coated ePTFE. However, specific incubation times for each coated graft should be selected and the long-term approach (graft culture) should also be attempted.
人成年内皮细胞通过酶解法从脂肪组织中获取。通过台盼蓝排斥试验以及透射和扫描电子显微镜确定细胞活力。通过光学显微镜、透射电子显微镜和扫描电子显微镜下的免疫细胞化学研究鉴定内皮细胞。分离出的细胞肌动蛋白和波形蛋白呈阳性，结蛋白呈阴性。因子VIII RA主要表达于细胞表面，偶尔在细胞质中可见。对UEA I和J15的反应较弱或无法检测到。将人内皮细胞接种到不同的非血管基质（玻璃、聚-L-赖氨酸、福尔马甲酸碳、纤连蛋白、聚四氟乙烯）上并使其贴壁1小时。扫描电子显微镜确定了表面特征，表明细胞在基质上牢固粘附。测试了不同的血管基质（预凝涤纶、白蛋白涤纶、Hemashield涤纶、Gelseal涤纶、ePTFE、纤连蛋白-ePTFE）。市售的涂层移植物在细胞粘附方面表现出定性和定量的差异。特别是，Gelseal涤纶提供了最好的定量结果，尽管观察到有很大的变异性。相比之下，纤连蛋白涂层的ePTFE给出了更可靠的结果和高铺展效率。短期内，涂层移植物似乎并不比纤连蛋白涂层的ePTFE具有更大优势。然而，应针对每种涂层移植物选择特定的孵育时间，并且也应尝试长期方法（移植物培养）。
