Use of freshly isolated capillary endothelial cells for the immediate establishment of a monolayer on a vascular graft at surgery.
Use of freshly isolated capillary endothelial cells for the immediate establishment of a monolayer on a vascular graft at surgery.
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使用新鲜分离的毛细血管内皮细胞在手术时在血管移植物上立即建立单层细胞。
作者：
B. Jarrell;Stuart K. Williams;G. Stokes;Hubbard Fa;R. Carabasi;E. Koolpe;Deborah A. Greener;K. Pratt;Moritz Mj;Radomski Js
血管移植物表面的内皮种植可能导致表面血栓形成较少。我们研究了使用源自人体脂肪的微血管内皮细胞进行接种的可行性。将人体脂肪用胶原酶处理 24 分钟、洗涤并在 Percoll 梯度分离中纯化。这产生了 1.25 +/- 0.45 X 10(6) 细胞/克脂肪。在等离子涂层涤纶上孵育 1 小时后，2.8 +/- 1.5 X 10(4) 个细胞仍牢固地粘附在表面上。当在 0 至 80 达因/cm2 的剪切应力下暴露于流动 2 小时时，50% 至 100% 的最初贴壁细胞保持贴壁。该数据的统计分析未能证明贴壁细胞数量与剪切速率之间存在密切关系。扫描电子显微镜显示内皮细胞处于附着于等离子体涂层涤纶的不同阶段。尽管大多数细胞仍然是圆形的并且仅局部附着在表面上，但一些细胞被最大限度地压平，形成细胞与细胞的接触。由于高细胞产量和牢固的粘附特性，我们得出结论，微血管内皮细胞可能为手术植入时移植物的汇合内皮细胞接种提供了可能性，而无需细胞培养。
Endothelial seeding of vascular graft surfaces may lead to a less thrombogenic surface. We examined the feasibility of using microvessel endothelial cells derived from human fat for seeding purposes. Human fat was treated with collagenase for 24 minutes, washed, and purified in a Percoll gradient separation. This yielded 1.25 +/- 0.45 X 10(6) cells/gm of fat. After a 1-hour incubation on plasma-coated Dacron, 2.8 +/- 1.5 X 10(4) cells remained firmly adherent to the surface. When exposed to flow for 2 hours at a shear stress of 0 to 80 dyne/cm2, between 50% and 100% of the initially adherent cells remained adherent. Statistical analysis of this data failed to demonstrate a strong relationship between the number of adherent cells and the shear rate. Scanning electron microscopy demonstrated endothelial cells in various stages of attachment to the plasma-coated Dacron. Although most cells were still round and only focally attached to the surface, some cells were maximally flattened, forming cell-to-cell contact. Because of the high cell yield and the firm adherence characteristics, we conclude that microvessel endothelial cells may offer the possibility for confluent endothelial cell seeding of a graft at the time of surgical implantation without the need for cell culture.