Specific induction of neuronal cells from bone marrow stromal cells and application for autologous transplantation

Abstract

Bone marrow stromal cells (MSCs) have the capability under specific conditions of differentiating into various cell types such as osteocytes, chondrocytes, and adipocytes.Here we demonstrate a highly efficient and specific induction of cells with neuronal characteristics, without glial differentiation, from both rat and human MSCs using gene transfection with Notch intracellular domain (NICD) and subsequent treatment with bFGF, forskolin, and ciliary neurotrophic factor.MSCs expressed markers related to neural stem cells after transfection with NICD, and subsequent trophic factor administration induced neuronal cells.Some of them showed voltage-gated fast sodium and delayed rectifier potassium currents and action potentials compatible with characteristics of functional neurons.Further treatment of the induced neuronal cells with glial cell line-derived neurotrophic factor (GDNF) increased the proportion of tyrosine hydroxylase-positive and dopamine-producing cells.Transplantation of these GDNF-treated cells showed improvement in apomorphine-induced rotational behavior and adjusting step and paw-reaching tests following intrastriatal implantation in a 6-hydroxy dopamine rat model of Parkinson disease.This study shows that a population of neuronal cells can be specifically generated from MSCs and that induced cells may allow for a neuroreconstructive approach.Nonstandard abbreviations used: anteroposterior (A/P); artificial cerebrospinal fluid (ACSF); bone marrow stromal cells (MSCs); brain-derived neurotrophic factor (BDNF); ciliary neurotrophic factor (CNTF); dopamine transporter (DAT); forskolin (FSK); galactocerebroside (GalC); glial cell line-derived neurotrophic factor (GDNF); glial fibrillary acidic protein (GFAP); hairy/enhancer of split-1 (Hes1); 6-hydroxy dopamine (6-OHDA); Janus kinase (JAK); lateral (L); microtubule-associated protein-2ab (MAP-2ab); nerve growth factor (NGF); neural stem cell (NSC); neuronal progenitor cell (NPC); NICD-transfected MSC (N-MSC); N-MSC administrated with the trophic factors (TF-MSC); Notch intracellular domain (NICD); 3-phosphoglycerate dehydrogenase (3-PGDH); signal transducer and activator of transcription 1 (STAT1); tetrodotoxin (TTX); TF-MSC administrated with GDNF (G-MSC); tyrosine hydroxylase (TH); ventral (V). Conflict of interest:The authors have declared that no conflict of interest exists.