Neuron-Like Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells

Abstract

PurposeMesenchymal stem cells (MSCs) are multipotent and give rise to distinctly differentiated cells from all three germ layers. Neuronal differentiation of MSC has great potential for cellular therapy. We examined whether the cluster of mechanically made, not neurosphere, could be differentiated into neuron-like cells by growth factors, such as epidermal growth factor (EGF), hepatocyte growth factor (HGF), and vascular endothelial growth factor (VEGF).Materials and MethodsBMSCs grown confluent were mechanically separated with cell scrapers and masses of separated cells were cultured to form cluster BMSCs. As described here cluster of BMSCs were differentiated into neuron-like cells by EGF, HGF, and VEGF. Differentiated cells were analyzed by means of phase-contrast inverted microscopy, reverse transcriptase-polymerase chain reaction (RT-PCR), immunofluorescence, and immunocytochemistry to identify the expression of neural specific markers.ResultsFor the group with growth factors, the shapes of neuron-like cells was observable a week later, and two weeks later, most cells were similar in shape to neuron-like cells. Particularly, in the group with chemical addition, various shapes of filament structures were seen among the cells. These culture conditions induced MSCs to exhibit a neural cell phenotype, expressing several neuro-glial specific markers.Conclusionbone marrow-derived mesenchymal stem cells (BMSCs) could be easily induced to form clusters using mechanical scraping, not neurospheres, which in turn could differentiate further into neuron-like cells and might open an attractive possibility for clinical cell therapy for neurodegenerative diseases. In the future, we consider that neuron-like cells differentiated from clusters of BMSCs are needed to be compared and analyzed on a physiological and molecular biological level with preexisting neuronal cells, and studies on the possibility of their transplantation and differentiation capability in animal models are further required.

Mesenchymal stem cells and conditioned medium avert enteric neuropathy and colon dysfunction in guinea pig TNBS-induced colitis

“…This can probably be attributed to prevention of neuronal loss rather than differentiation of MSCs into enteric neurons. Several studies have described the in vitro differentiation of MSCs into cells of the neuronal and glial lineage (3,88), but this occurs only when MSCs are cultured under particular experimental conditions as discussed above. The neuroprotective effects of MSC and CM treatments found in our study correlate with previous reports in animal models of CNS disorders (37,39), in which MSCs prevented axonal damage and promoted nerve fiber regeneration through the production of trophic factors.…”

“…MSCs play a major role in tissue regeneration, repair, and homeostasis with capacity for self-renewal and differentiation into cells of mesodermal lineage such as bone, cartilage, muscle, and fat (4,20), as well as cells of neuronal and glial lineage when cultured under particular experimental conditions (3,88). These multipotent cells can be derived from many adult tissues, including bone marrow (BM), adipose tissue, umbilical cord, and placenta (71).…”

Transplantation of bone marrow mesenchymal stem cells pretreated with valproic acid in rats with an acute spinal cord injury

“…All 3 groups of cells were plated in 6-well tissue culture plates and growth medium was replaced. Cells were then fed by completely replacing the medium with fresh BMSC osteogenic/adipogenic differentiation medium (Cyagen Biosciences Inc., California, USA) or growth medium supplemented with 10 ng/mL b-fibroblast growth factor (bFGF, Gibco) and 10 ng/mL epidermal growth factor (EGF, Gibco) (12). After differentiation, cells were fixed and subjected to Alizarin red staining, Oil red O staining, and immunofluorescence staining.…”

“…Hind limb locomotion was evaluated using the Basso, Beattie, and Bresnahan (BBB) locomotor rating scale on days 1, 3, 7, 14, and 28 after SCI (12). During the evaluation, rats were placed in an open field and observed for 5 minutes at each time point by 2 observers who were blind to the experimental protocol.…”

Study of the involvement of allogeneic MSCs in bone formation using the model of transgenic mice

“…5 Under certain in vitro conditions MSCs are capable of differentiating into osteoblasts, adipose cells, cartilage cells, skeletal muscle cells and even into neuron-like cells. 6,12 MSCs lack expression of the major histocompatibility complex I and of costimulatory molecules like CD40, CD80, and CD86, which makes them largely nonimmunogenic. 13 Furthermore, these cells are capable of suppressing the immune system via still unknown mechanisms, thus, the treatment with such allogeneic cells is largely immunoinert.…”