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.

Updates in the pathophysiological mechanisms of Parkinson’s disease: Emerging role of bone marrow mesenchymal stem cells

“…Brain tissue sections examination indicated that single infusion with BM-MSCs resulted in intact histological structure of the striatum. This finding coincides with Dezawa et al [84] who reported that nerve system recovery after BM-MSCs transplantation could be related to their secretion of neurotrophic factors that restore the function of nervous system, promotion of local angiogenesis and vascular reconstruction and neuronal regeneration through promotion of autologous neuronal regeneration and differentiation of transplanted cells into neural cells.…”

Ex vivo expansion and pluripotential differentiation of cryopreserved human bone marrow mesenchymal stem cells

“…For neurogenesis, various inducers, alone or in combination, are involved in neurocytic differentiation in vitro of MSCs. These include retinoic acid (Cho et al, 2005), DMSO and BHA (Chu et al, 2004;Lu et al, 2004), GM1 and bFGF (Dezawa et al, 2004;Zhang et al, 2004b;Munoz-Elias et al, 2003), and 2-mercaptoethanol (2-ME) (Guo et al, 2001). It was suggested that cAMP signaling pathway should be involved in neurocytic differentiation of MSCs (Suon et al, 2004;Jori et al, 2005), but it could not rule out the synergistic effect of other signaling pathways on neurogenesis of hMSCs, in regard to neural differentiation of embryonic stem cells (Kleber et al, 2005;Bainter et al, 2001;Otero et al, 2004).…”

Bone Marrow Mesenchymal Cells: How Do They Contribute to Tissue Repair and Are They Really Stem Cells?

“…The MSCinducible cell types include endothelial cells (Oswald et al 2004), cardiac muscle cells (Makino et al 1999), skeletal muscle cells (Dezawa et al 2005), hepatocytes (Oyagi et al 2006;Prindull and Zipori 2004;Snykers et al 2009), neuronal cells (Dezawa et al 2004;Wislet-Gendebien et al 2005), peripheral glial cells (Dezawa et al 2001) and insulin-producing cells (Phinney and Prockop 2007) and epithelial cell (Spees et al 2003) (Fig. 1).…”

“…Granted that MSCs are a heterogeneous cell population, MSCs as a whole show differentiation not only into the same mesodermal lineage cells of osteocytes, chondrocytes, and adipocytes, but also into other lineages such as into neuronal cells (ectoderm) and hepatocytes (endoderm) (Dezawa et al 2001(Dezawa et al , 2004(Dezawa et al , 2005Pittenger et al 1999). For this reason, MSCs are considered pluripotent cells.…”

“…For example, HSCs generate red blood cells and white blood cells, and neural stem cells generate neurons and glial cells (Gage 2000;Weissman and Shizuru 2008). MSCs differ from these somatic stem cells in that they differentiate not only into the same mesodermal-lineage, such as bone, cartilage, and adipocytes, but also into other lineages of ectodermal and endodermal cells (Dezawa et al 2004(Dezawa et al , 2005Pittenger et al 1999;Snykers et al 2009). In this manner, MSCs can generate cells representative of all three germ layers, and are thus thought to be pluripotent cells.…”