Simultaneous Measurement of VSMCs Metabolic Activity and Proliferation with WST-1 and Cell Proliferation ELISA(BrdU)

摘要

In laboratory evaluation of cell proliferation and metabolic activity the WST-1 assay is more sensitive and more convenient than the MTT assay. However, WST-1 is not metabolized by all cell types. To test wether WST-1 can be metabilized by the cultured vascular smooth muscle cells(VSMCs) derived from aortas of New Zealand white rabbits, we developed a combination of WST-1 assay and the Cell Proliferation ELISA, BrdU for evaluation of proliferation, metabolic activity and DNA synthesis of VSMCs. The WST-1 assay was carried out in a 96-well microtiterplate (MTP)in the presence of various concentrations of fetal calf serum (FCS). Cell Proliferation ELISA, BrdU was employed simultaneously in the same MTP to measure DNA synthesis. We found that VSMCs metabolic activity and proliferation increased in a dose-dependent manner in the presence of FCS (p<0.05). These data indicate that WST-1 is metabolized and can be used as a new parameter for measurement of VSMCs proliferation and metabolic activity. The WST-1 in combination with Cell Proliferation ELISA, BrdU assay is a useful tool to evaluate the specific effect of a given factor or drug on either cellular DNA syntheses or cellular metabolic activity or both.