Differential expression of microRNA-125b in the neuronal differentiation of adipose-derived Flk1~+ mesenchymal stem cells / 中国组织工程研究

ABSTRACT

BACKGROUND:The differentiation of mesenchymal stem cells(MSCs)is strictly regulated by both genetic and epigenetic Factors .Emerging evidences have demonstrated that microRNA also plays an important role in this process.OBJECTIVE:To explore the differential expression of microRNA-125b during neuronal dliferentiation of Flk1~+ adipose-derived MSCs(AD-MSCs).MATERIALS:The fat samples were provided bv healthy female volunteers aged 15-35 years and recruited from the Plastic Surgery Hospital affiliated to the Chinese Academy of Medical Sciences.METHODS:Adult adipose tissues were obtained from healthy voluntears with age of 15-35 years .Using adherence method,Flk1~+ MSCs were obtained and the 3~(rd) passage cells were taken in the experiment.Cultured in neuronalinduction medium.these MSC were induced to differentiate towards neuronal lineage.The expression of microRNA-125b was examined at days 0,4,8 and 12.To explore its role in neuronal differentiation,we need to change its expression.RT-PCR and Taqman real-time PCR were carried out to explore the difierentially expression of microRNA-125b during neuronal differentiation of AD-MSCs.The effect of inhibitor on the expression of microRNA-125b was detected.RESULTS AND CONCLUTION:①The Flk-1~+MSCs were successfully induced into neuronal differentiation and displayed typical morphological changes 12 days after induction:Most cells retracted their cytoplasm ,fomling spherical cell body and emitted cellular protrusions.RT-PCR and immunocytochemistry studies confirmed their phenotype with expression of known neuronal cell markers including neurofilament,glial fibrillary acidic protein.②The expression of microRNA-125b was significant up-regulated during neuronal differentiation .Results of RT-PCR and Taqman real-time PCR were concordarlce with that of microRNA chip technology.③Inhibitor could down-regulate microRNA-125b.The results implied that microRNA-125b may play an important role in neuronal differentiation.