NRSF silencing induces neuronal differentiation of human mesenchymal stem cells

• Published in: Experimental cell research Vol. 314; no. 11; pp. 2257 - 2265
• Main Authors: Yang, Yinxiang, Li, Yanhua, Lv, Yang, Zhang, Sainan, Chen, Lin, Bai, Cixian, Nan, Xue, Yue, Wen, Pei, Xuetao
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.07.2008; Elsevier BV
• Subjects: Biomarkers - metabolism; Bone marrow; Bone Marrow Cells - cytology; Bone Marrow Cells - physiology; Cell Differentiation - physiology; Cell Lineage; Cells, Cultured; Cellular biology; Gene expression; Human mesenchymal stem cells; Humans; Ion Channels - metabolism; Lentivirus; Lentivirus - genetics; Lentivirus - metabolism; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - physiology; Neuronal differentiation; Neuronal restrictive silencing factor (NRSF)/repressor element-1 (RE-1) silencing transcription factor (REST); Neurons; Neurons - cytology; Neurons - physiology; Patch-Clamp Techniques; Repressor Proteins - genetics; Repressor Proteins - metabolism; RNA interference (RNAi); RNA, Small Interfering - genetics; RNA, Small Interfering - metabolism; Stem cells; Neuronal differentiation; Neuronal restrictive silencing factor (NRSF)/repressor element-1 (RE-1) silencing transcription factor (REST); RNA interference (RNAi); Human mesenchymal stem cells
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1016/j.yexcr.2008.04.008

Mesenchymal stem cells (MSCs) are multipotent cells that have the potential to differentiate into the neuronal cell lineage. Here, we describe the highly efficient and specific induction of cells with neuronal characteristics, without glial differentiation, from human bone marrow-derived mesenchymal stem cells by NRSF silencing. Cells that have the characteristics of MSCs were obtained from human bone marrow. Lentiviral vectors were used to deliver small interference NRSF RNA (siNRSF) into MSCs. After being infected with lentivirus containing siNRSF, MSCs were successfully induced to differentiate into neuronal cells, which exhibited neuron-like morphology and formed nissl bodies. These differentiated cells expressed multiple neuron-specific genes including brain-derived neurotrophic factor (BDNF), neurogenin 1 (NGN1), neuron-specific enolase (NSE), synaptophysin (SYP), and neuron-specific growth-associated protein (SCG10), as well as expressing mature neuronal marker proteins, such as β-tubulin III, NSE, microtubule-associated protein type 2 (MAP-2), and neurofilament-200 (NF-200), yet did not express the glial markers glial fibrillary acidic protein (GFAP) and oligodendrocyte transcription factor 2 (Olig2), as verified by immunofluorescence staining. The whole cell patch-clamp technique recorded TTX-sensitive Na + currents and action potential from these differentiated cells. Thus, our results demonstrate that NRSF silencing can activate some neuronal genes and induce neuronal differentiation of mesenchymal stem cells.