Culture and identification of rat basilar artery smooth muscle cells / 中国药理学通报

Aim To investigate the method of cell culture for smooth muscle cells from rat cerebral basilar artery and understand cells growth and biological characteristics.Methods The explant attached method was applied for cell culture of rat basilar artery smooth muscle cells(BASMCs).The cultured BASMCs were identified by immunocytochemical staining.The activities of cells were indicated by the dynamic changes of intracellular calcium concentration observed by RF-5 000 fluorospectro-photometer.Results BASMCs grew out of tissue blocks by 5 days,reached confluency could be subcultured after 2 weeks.Cultured cells were identified by intensely positive immunocytochemical staining to smooth muscle actin-specific.Introduction of calcium channel agonists induced significant increase in Fura-2 fluorescence ratio(F340/F380)and cells were in good condiction.Conclusion Explant attached method is simple,efficient and economic.It provides an ideal cell model for the study of pathogenesis of the cerebral vascular diseases.

The interference of Cd~(2+) with Fura-2 fluorescence in measurement of free cytoplasmic Ca~(2+) / 中国药理学通报

Aim To investigate the blockade of cadmium on store operated calcium channels and the fluorescence interference of cadmium with Fura-2.Methods PC12 cells were used to determine the intracellular calcium concentration [Ca~(2+)]i indicated by change in Fura-2 fluorescence ratio(F_(340)/F_(380)).Results Introduction of cadmium induced a significant increase in Fura-2 fluorescence ratio following thapsigargin-evoked calcium entry;Besides,cadmium gave rise to a remarkable elevation in Fura-2 fluorescence ratio following breakdown of the plasma membrane by triton,a detergent.The Fura-2 fluorescence was increased in the presence of cadmium and Fura-2/AM,simultaneously in the absence of PC12 cells.Conclusion Cadmium can interfere with the Fura-2 fluorescence.