Components of subendothelial aorta basement membrane. Immunohistochemical localization and role in cell attachment

Cryosections of fetal and adult bovine aorta were stained with purified, cross-absorbed antibodies against various connective tissue components. The antibodies to the basement membrane components, laminin, heparan sulfate proteoglycan, and type IV collagen, gave a sharp reaction in the subendothelial layer. Antibodies against type III procollagen showed a broad endothelial staining, and staining was also seen in the media layer. A similar staining reaction was seen with antibodies against fibronectin. Bovine fetal aortic endothelial (BAE) cells were isolated and cultured in vitro. The cells became stained by the indirect immunofluorescence method with antibodies against laminin and heparan sulfate proteoglycan and also with antibodies against types III and IV collagen and type I procollagen, as in previously reported experiments. The attachment properties of endothelial cells to the same extracellular matrix components were also studied. BAE cells became attached most readily to surfaces coated with fibronectin or type III or type IV collagen. Laminin and collagen types I and V served as less effective substrates. Attachment to heparan sulfate proteoglycan was slowest of the tested components. The results of the study demonstrate that the BAE cells are associated with basement membranes in vivo. The BAE cells in culture produced interstitial connective tissue components in addition to basement membrane components and showed no clear specific preference in their attachment to any of these.