A New Multistep Induction Protocol for the Transdifferentiation of Bone marrow Stromal Stem Cells into GABAergic Neuron-Like Cells

A New Multistep Induction Protocol for the Transdifferentiation of Bone marrow Stromal Stem Cells into GABAergic Neuron-Like Cells

A New Multistep Induction Protocol for the Transdifferentiation of Bone marrow Stromal Stem Cells into GABAergic Neuron-Like Cells

ABSTRACT

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. Iran

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