EXPRESSION AND ELECTROPHYSIOLOGICAL CHARACTERISTICS OF L-TYPE CALCIUM CHANNEL IN RAT BONE MESENCHYMAL STEM CELLS / 解剖学报

ABSTRACT

Objective To explore the electrophysiological characteristics and the expression of mRNA and protein of L-type calcium channel in rat bone mesenchymal stem cells(MSCs). Methods MSCs were isolated,cultured and purified.RT-PCR was used to detect the mRNA expression of ?1C,?1D,?1H,?1S.The protein expression of L-type calcium channel(?1C) was testified by immunohistochamical.Ion currents were recorded in MSCs using whole-cell patch clamp technques.Results CD29,CD44,CD106 expressed in about 93% MSCs and CD14,CD34,CD45 expressed negatively.A high expression of mRNA in ?1C was detected by RT-PCR but no expressions were observed in ?1D,?1G,?1H,?1S.Immunofluorescent double labeling showed an expression of ?1C subunits in MSCs.Moreover,inward currents were recorded in 16 of 36 cells using whole-cell patch clamp techniques.The currents were activated around-30?mV and peaked at 0 to 10?mV and were blocked by nifedipine(10??mol/L).These cells had larger currents with Ba~(2+)(10?mmol/L) in bath solution than with Ca~(2+)(2?mmol/L).Conclusion The results indicated that adult rat MSCs expresse functional L-type calcium channels.It is possible that this channel plays a role on the proliferation and differentiation of MSCs.