Radomski J S, Jarrell B E, Williams S K, Koolpe E A, Greener D A, Carabasi R A
J Surg Res. 1987 Feb;42(2):133-40. doi: 10.1016/0022-4804(87)90110-7.
Successful endothelialization of vascular grafts by seeding with endothelial cells (EC) at implantation is related to the number of EC which initially adhere to the graft. Using an in vitro system we examined the initial adherence of EC from human perinephric fat capillaries to woven Dacron that was either unmodified or precoated with several substrates. We studied capillary EC because they have not been investigated as a source of EC for graft seeding, although transinterstitial capillary ingrowth is one possible mechanism for spontaneous graft endothelialization. EC were isolated using collagenase and characterized morphologically and functionally including positive factor VIII-related antigen staining. EC were studied at three phases in culture: (A) primary EC with no subcultivations (EC-0); (B) EC after two subcultivations with trypsin (EC-2); and (C) EC after 10 subcultivations with trypsin (EC-10). EC were seeded onto graft material at a density of 10(5) cells/cm2 (100% confluence) and examined for cell counts and morphology after one day in culture by light and electron microscopy. Results are as follows: (table; see text) The conclusions are: (1) All capillary EC demonstrated adherence to Dacron, but this initial adherence was strongly influenced by graft pretreatment with collagen or plasma. (2) Serially subcultivated EC (EC-2 and EC-10) had significantly higher initial adherence to pretreated Dacron compared to the primary cells (EC-0) (P less than .05). This suggests that briefly cultured and subcultivated EC have superior initial adherence characteristics to treated dacron compared to primary EC with no subcultivations. (3) Fat capillary EC are easily procured and cultured and provide a rich source of human EC for endothelializing vascular prostheses.
在植入时通过接种内皮细胞(EC)使血管移植物成功内皮化,这与最初黏附于移植物的EC数量有关。我们利用体外系统,研究了来自人肾周脂肪毛细血管的EC对未修饰或预先包被了几种基质的编织涤纶的初始黏附情况。我们研究毛细血管EC,是因为尽管跨间质毛细血管向内生长是移植物自发内皮化的一种可能机制,但尚未将其作为移植物接种的EC来源进行研究。使用胶原酶分离EC,并对其进行形态学和功能特性鉴定,包括因子VIII相关抗原染色阳性。在培养的三个阶段研究EC:(A)未传代的原代EC(EC-0);(B)用胰蛋白酶传代两次后的EC(EC-2);(C)用胰蛋白酶传代10次后的EC(EC-10)。将EC以10⁵个细胞/cm²的密度接种到移植物材料上(100%汇合),并在培养一天后通过光学和电子显微镜检查细胞计数和形态。结果如下:(表格;见正文)结论如下:(1)所有毛细血管EC均表现出对涤纶的黏附,但这种初始黏附受到胶原或血浆预处理移植物的强烈影响。(2)与原代细胞(EC-0)相比,连续传代培养的EC(EC-2和EC-10)对预处理涤纶的初始黏附显著更高(P<0.05)。这表明,与未传代的原代EC相比,短暂培养和传代的EC对处理后的涤纶具有更好的初始黏附特性。(3)脂肪毛细血管EC易于获取和培养,为血管假体的内皮化提供了丰富的人EC来源。