Development of a Rotation Device for Microvascular Endothelial Cell Seeding

A rotation device (RD) specifically designed to achieve sterile endothelial cell (EC) seeding of vascular grafts has been developed. The basic characteristics of the RD include: small dimensions , fully autoclavable components, and perfectly sealed graft holders. These features make it possible to maintain sterility during all the steps of EC seeding. This was documented by negativity of all bacteriological assays performed . Moreover , the RD can simultaneously support three vascular grafts with different lengths (20 , 40, and 60 cm) and diameters (4-8 mm). EC seeding is performed in the cli matized chamber (37 °C; 5 % C02) with constant rotation (0.1 -3 rpm). The rotation cycle can be completed automatically. The practical efficacy of the RD was investigated by seeding 2 x 105/cm2 of human microvascular EC on 20 cm length , 4 mm internal diameter (ID) fi bronectin-coated polytetrafluoroethylene (ePTFE) grafts for 24 and 48 hours respectively . Further, the effect of a highly viscous plasma expander , i.e . , haemagel , on cell retention was also evaluated. Results were not as favorable as expected. However , it should be emphasized that after 48 hours of eel! incubation by using the RD, 42 % of the initially seeded EC were still present and approximately 15 % were fully spread over the graft surface. Moreover, the 10 minute perfusion with haemagel did not decrease the number of adherent microvascular EC.