TY - JOUR T1 - A New Multistep Induction Protocol for the Transdifferentiation of Bone marrow Stromal Stem Cells into GABAergic Neuron-Like Cells TT - پروتکل چند مرحله ای جدید در تمایز سلولهای استرومایی مغز استخوان به نورونهای شبه گاباارژیک JF - انستیتو-پاستور-ایران JO - انستیتو-پاستور-ایران VL - 17 IS - 1 UR - http://ibj.pasteur.ac.ir/article-1-812-en.html Y1 - 2013 SP - 8 EP - 14 KW - Cell therapy KW - GABAergic neurons KW - Creatine KW - Bone marrow stem cell N2 - 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. M3 10.6091/IBJ.1112.2012 ER -