Establishment Of An In-Vitro Cell Model Of Cochlear Microcirculation And Potassium Channel TREK-1 Expression

Objective To establish an in-vitro cell model of cochlear microcirculation, investigate expression of mechanosensitive potassium channel TREK-1 in this in-vitro cell model,explore the function and significance of TREK-1 in regulation of local blood flow in cochlea.Methods Endothelial cells were primary cultured from stria vascularis of rat cochlea.After passage and purification,the immunocytochemistry was employed to classify cell types by examining the marked antigen of endothelial cells.The expression of TREK-1 was detected by reverse transcription-polymerase chain reaction(RT-PCR) and immunofluorescence in these cultured cells.Results 1,After passage and purification,the cultured cells showed typical "flagstone" appearance as endothelial cells.According to immunocytochemiscal detection of factorâ…§, we observed that the cultured cells show positive reaction.2,The TREK-1 cDNA was specifically amplified by RT-PCR,the immunofluorescence which was performed to detect TREK-1 protein expression showed positive reaction. Conclusion The method used in this study can obtain endothelial cells of the stria vascularis,which can be used as an in vitro model for cochlear microcirculation.The expression of mechanosensitive potassium channel TREK-1 was confirmed in endothelial cells of stria vascularis,suggesting TREK-1 may be involved in regulation of local blood flow in cochlea.