a new multistep induction protocol for the transdifferentiation of bone marrow stromal stem cells into gabaergic neuron-like cells

background: bone marrow stromal stem cells (bmsc) are appropriate source of multipotent stem cells that are ideally suited for use in various cell-based therapies. it can be differentiated into neuronal-like cells under appropriate conditions. this study examined the effectiveness of co-stimulation of creatine and retinoic acid in increasing the differentiation of bmsc into gabaergic neuron-like cells (gnlc). methods: bmsc isolated from the femurs and tibias of adult rats were cultured in dmem/f12 medium supplemented with 10% fbs, pre-induced using β-mercaptoethanol (βme) and induced using retinoic acid (ra) and creatine. immunostaining of neurofilament 200 kda, neurofilament 160 kda, nestin, fibronectin, gamma-amino butyric acid (gaba) and glutamic acid decarboxylase (gad) 65/67 were used to evaluate the transdifferentiation of bmsc into gnlc and to evaluate the effectiveness of pre-induction and induction assays. the expression of genes that encode fibronectin, octamer-binding transcription factor 4 (oct-4), gad 65/67 and the vesicular gaba transporter was examined in bmsc and gnlc by using rt-pcr assays during transdifferentiation of bmsc into glnc. results: co-stimulation with ra and creatine during the induction stage doubled the rates of gabaergic differentiation compared with induction using creatine alone, resulting in a 71.6% yield for glnc. rt-pcr showed no expression of oct-4 and fibronectin after the induction stage. conclusion: the results of this study showed that the application of βme, ra, and creatine induced the transdifferentiation of bmsc into glnc.