Differentiation of Equine Mesenchymal Stromal Cells into Cells of Neural Lineage: Potential for Clinical Applications

Abstract

Mesenchymal stromal cells (MSCs) are able to differentiate into extramesodermal lineages, including neurons. Positive outcomes were obtained after transplantation of neurally induced MSCs in laboratory animals after nerve injury, but this is unknown in horses. Our objectives were to test the ability of equine MSCs to differentiate into cells of neural lineage in vitro, to assess differences in morphology and lineage-specific protein expression, and to investigate if horse age and cell passage number affected the ability to achieve differentiation. Bone marrow-derived MSCs were obtained from young and adult horses. Following demonstration of stemness, MSCs were neurally induced and microscopically assessed at different time points. Results showed that commercially available nitrogen-coated tissue culture plates supported proliferation and differentiation. Morphological changes were immediate and all the cells displayed a neural crest-like cell phenotype. Expression of neural progenitor proteins, was assessed via western blot or immunofluorescence. In our study, MSCs generated from young and middle-aged horses did not show differences in their ability to undergo differentiation. The effect of cell passage number, however, is inconsistent and further experiments are needed. Ongoing work is aimed at transdifferentiating these cells into Schwann cells for transplantation into a peripheral nerve injury model in horses.

A Novel Model for Acute Peripheral Nerve Injury in the Horse and Evaluation of the Effect of Mesenchymal Stromal Cells Applied In Situ on Nerve Regeneration: A Preliminary Study

“…Western blot and immunofluorescence analyses revealed the expression of the Schwann cell markers S-100b and GFAP in SLCs. Undifferentiated MSCs also expressed β 3 and GFAP, as previously reported by our group ( 23 ), but not S-100b. Expression of β 3 and GFAP suggests that EBM-MSCs are capable of differentiating into cells of extra-mesodermal lineages ( 31 ).…”

“…These studies are necessary before EBM-MSCs can be used in veterinary medicine to enhance regeneration of damaged peripheral nerves. We have recently reported that EBM-MSCs are able to display morphological and protein characteristics of neural progenitors ( 23 ). In this preliminary study, we evaluated the ability of EBM-MSCs to differentiate into SLCs after chemical induction, in vitro .…”

Promoting effect of small molecules in cardiomyogenic and neurogenic differentiation of rat bone marrow-derived mesenchymal stem cells

“…As mentioned earlier, several studies have described different methods of chemical induction for neuronal differentiation; however, results from different studies are contradictory and hence inconclusive and mostly showed less number of interconnecting neurons. 29 , 38 In our study, MSCs were morphologically differentiated into neuronal-like cells after second induction. Untreated MSCs exhibited fibroblast-like cells in the control group.…”

“…Many agents such as cytokines, growth factors, neurotrophins, and small molecules have been shown to promote neuronal and cardiac cell differentiation both in vivo and in vitro. 27 – 29 Woodbury et al 4 showed the positive effect of some antioxidants, dimethyl sulfoxide, butylated hydroxyanisole, and β-mercaptoethanol (BME), on the differentiation of BM-MSCs into neuronal cells. Among all the inducing agents, BME and 5-azacytidine appeared to be well-known agents for stem cell differentiation into neuronal cells and cardiomyocytes, respectively.…”

Bone Marrow-Derived, Neural-Like Cells Have the Characteristics of Neurons to Protect the Peripheral Nerve in Microenvironment

“…Bone marrow mesenchymal stem cells (BM-MSCs) injected into the site of nerve injury can differentiate into Schwann-like cells and support nerve regeneration [ 12 – 14 ]. It has been postulated that BM-MSCs release nerve growth factor that stimulates Schwann cell proliferation and enhances sciatic nerve regeneration [ 15 ].…”