ABILITY OF VARIOUS INSERTS TO PROMOTE ENDOTHELIUM CELL-CULTURE FOR THE ESTABLISHMENT OF COCULTURE MODELS

To select an insert suitable for human umbilical vein endothelial cell (HUVEC) culture, we compared several available inserts of 0.2 to 0.45 mu m porosity: Cellagen (ICN), Transwell-COL (Costar), Millicell-HA a nd CM (Millipore), Anopore (Nunc), Cyclopore (Falcon) in comparison wi th a control surface (Thermanox). The requirements were: (i) to promot e attachment, adhesion and proliferation of HUVEC (judged by [H-3]thym idine incorporation into DNA at days 1, 3, 7); (ii) to allow HUVEC vis ualization by inverted, fluorescence microscopy for uptake of DiI-Ac-L DL and scanning electron microscopy, performed at day 9 after seeding. Because Transwell and Cellagen are collagen precoated and CM has to b e coated for cell culture, we performed collagen coating (types I + II I or IV) for non-pretreated inserts for the purpose of comparison. Our preferences comprise Transwell-COL, Cyclopore not coated or coated (w hatever the collagen type), and Cellagen. However, on a quality/price ratio criterion, Cyclopore, even uncoated, is the insert of choice. Th e HA, CM and Anopore inserts, even coated, do not allow HUVEC growth b ut do not alter positive uptake of acetylated LDL.