Abstract
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/cm 2 (100% confluence) and examined for cell counts and morphology after one day in culture by light and electron microscopy. Results are as follows: Dacron pretreatment EC-O (%) EC-2 (%) EC-10 (%) Untreated 37.4 + 1.4 43.3 + 2.2 44.2 + 5.3 Collagen I/III 29.5 + 2.6 58.7 + 2.4 46.6 + 7.9 Platelet-rich plasma 52.9 + 0.4 64.6 + 3.3 62.8 + 7.1 Collagen-plus plasma 34.3 + 3.2 47.1 + 1.1 65.8 + 3.6 Percentage of capillary EC adherent to dacron following seeding at confluent density (100% = 10 5 cells/cm 2) + SEM 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 < .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.
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