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Brain Res Bull. 2013 Oct;99:84-94. doi: 10.1016/j.brainresbull.2013.10.005. Epub 2013 Oct 19.

Over-expression of Mash1 improves the GABAergic differentiation of bone marrow mesenchymal stem cells in vitro.

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1
Department of Neurosurgery, Xijing Institute of Clinical Neuroscience, Fourth Military Medical University, No.17 Chang-le West Road, Xi'an 710032, China; Department of Neurosurgery, Qingdao 401 Hospital of PLA, No. 22 Minjiang Road, Qingdao 266071, China.

Abstract

Bone marrow mesenchymal stem cells (BMSCs) have been shown to be a promising cell type for the study of neuronal differentiation; however, few attempts had been made to differentiate these cells into inhibitory gamma-aminobutyric acid (GABA)ergic neurons. In this study, we over-expressed mammalian achaete-scute homologue-1 (Mash1), a basic helix-loop-helix (bHLH) transcription factor, in Sprague-Dawley rat BMSCs via lentiviral vectors, and then induced neuronal differentiation of these cells using conditioned medium. Our Western blot results show that, under conditions of differentiation, Mash1-overexpressing BMSCs exhibit an increased expression of neuronal markers and a greater degree of neuronal morphology compared to control, non-Mash1-overexpressing cells. Using immunocytochemistry, we observed increased expression of glutamic acid decarboxylase 67 (GAD67), as well as neuron-specific nuclear protein (NeuN) and β3-tubulin, in Mash1-overexpressing BMSCs compared to control cells. Moreover, we also found the differentiated cells showed representative traces of action potentials in electrophysiological characterization. In conclusion, our study demonstrated that over-expression of Mash1 can improve GABAergic differentiation of BMSCs in vitro.

KEYWORDS:

BMSCs; Bone marrow mesenchymal stem cell; Differentiation; GABA; GAD67; Mash1; NeuN; bHLH; basic helix-loop-helix; bone marrow mesenchymal stem cells; gamma-aminobutyric acid; glutamic acid decarboxylase 67; mammalian achaete-scute homologue-1; neuron-specific nuclear protein

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