Expressions of ionic channels of bone marrow mesenchymal stem cells and their significance / 吉林大学学报(医学版)

ABSTRACT

Objective To investigate the expression of ionic channels of the rat bone marrow mesenchymal stem cells(BMSCs) and provide the basis for the change of ionic channels in the process of differentiation of stem cells.Methods The full marrow from Wistar rats was isolated by Percoll centrifugation and passaged repeatedly.The membrane currents of BMSCs were recorded with the whole-cell patch-clamp technique.The current amplitude of pathway,peal value and time course of activation and deactivation,selectivity of ions pathway,I-V curve were recorded.The currents were identified and analyzed by using various channel blockers.Results The cells began to stick in 24 h and arranged in whirlpool shape after passenger culture.They were positive for CD71,CD44,and negative for CD34,CD45 by flow cytometry,which improved that the cultured cells were BMSCs.The functional ion channels were characterized in cultured rat BMSCs with whole-cell patch clamp and reverse transcription polymerase chain reaction(RT-PCR) techniques.Three types of outward currents were found in BMSCs,including a delayed rectifier K+ current(IKDR),in addition,tetrodotoxin-sensitive sodium current(INa.TTX) and nifedipine-sensitive L-type Ca2+ current(ICa.L) were detected in 24% and 25% BMSCs.Moreover,RT-PCR result revealed the molecular evidence of mRNA for the functional ionic currents,including SCN5A,Kv4.3 and CACNA1C.Conclusion The currents are consistent with the characteristics of excitable cell membrane.