Improved growth of cultured brain microvascular endothelial cells on glasscoated with a biological matrix

An improved method for culturing primary rat brain capillary endothelial ce lls on glass has been developed, using a corneal extracellular matrix coat. Since the collagen-coated plastic attachment surface conventionally used f or primary cultures of brain microvascular endothelium gives a high level o f background fluorescence in microfluorimetric studies, an alternative atta chment surface was tested involving no plastic element. Five substrate comb inations were examined and a new combination of glass and corneal endotheli al extracellular matrix coat was found to provide excellent cell adhesion; culture growth and purity. Other established substrata combinations tested for comparison, either involved plastic, or used glass with collagen or car bodiimide and collagen coating but the last two gave poor endothelial cell adhesion and growth. Our method using this new attachment surface combinati on results in stable and pure endothelial cultures, as verified by immunocy tochemistry, which are suitable for fluorimetric investigations. (C) 1999 E lsevier Science Ltd. All rights reserved.