RESUMEN
Objective To determine the expression pattern of carbohydrate chains in two kinds of microvascular endothelial cells (MVECs). Methods Rat jejunal mucosal MVECs were thawed, and lung tissues were removed from specific pathogen free piglet of 3 days old to isolate and culture porcine pulmonary MVECs by collagerase digestion and differential attachment. By lectin cytochemistry, staining of 8 lectins including concanavalin A (Con A), phaseolus vulgaris erythroagglutinin (PHA-E), ricinus communis agglutinin I (RCA-I), lycopersicon esculentum lectin (LEL), sambucus nigra lectin (SNA), ulex europaeus agglutinin I (UEA-I), wheat germ agglutinin (WGA) and dolichos biflorus agglutinin (DBA) was detected in rat jejunal mucosal and porcine pulmonary MVECs. Results In rat jejunal mucosal MVECs, strong positve staining was present for Con A, WGA and LEL, medium one for PHA-E N SNA and RCA-I, weak one for DBA, and negative staining for UEA-I. In porcine pulmonary MVECs, strong positive staining was present for Con A and PHA-E, medium one in RCA-I, weak one for LEL and SNA, and negative staining for UEA-I, WGA and DBA. Conclusion The carbohydrate patterns in two kinds of MVECs display significant heterogeneity. Both rat jejunal mucosal and porcine pulmonary MVECs express mannose, galactose, 1, 3-N-acetylglucosamine and sialic acid at different levels. N-acetylglucosamine and N-acetylgalactosamine are detected in the former but not in the latter, and fucose do not in both MVECs.
RESUMEN
A modified protocol for quick and economic treatment of cultured human umbilical vein endothelial cells has been established, which result in high viability of the cells to allow better performance in patch-clamp studies. The electrophysiological properties of Ca (2+)-activated K(+) (KCa) channel of the cultured cells were investigated with a cell-attached configuration. With this modified treatment method, the cultured cells appear fusiform in shape under microscope, and KCa channel currents could be detected readily, suggesting their eligibility for patch-clamp studies.