The Influence of Cerebrospinal Fluid Accompanied by Retinoic Acid on Differentiation of Bone Marrow Mesenchymal stem Cells into Neuron-like Cells in vitro

Cerebrospinal fluid (CSF) has a broad range of molecules and neurotrophic factors which are essential for neurogenesis. Bone marrow mesenchymal stem cells (BMSCs) are multipotent stem cells that can differentiate into the cells with neural-like phenotype under the induction of appropriate growth factors. According to the significant role of Retinoic acid (RA) in neurogenesis, The aim of this study is differentiation of BMSCs into neuron-like cells using CSF, RA and combinative form of CSF and RA.
Material and Rat BMSCs were isolated and characterized. The CSF was prepared from cisterna magna of 19 days Wistar rat embryos. Then BMSCs were induced by 5% CSF (CSF group), 10-6 µM Retinoic acid (RA group) and CSF plus RA (CSR group) for 12 days. Morphology of differentiated cells was examined by inverted microscope and axonal outgrowth was measured using the image J software. In addition, the expression of neural-specific markers (Nestin and MAP-2) was examined by Immunocytochemistry.
The specific - neuronal morphology was observed in differentiated cells. The maximum axon length was seen in CSR group on 12th day of induction. The results of immunocytochemistry showed that the neuroprogenitor marker (Nestin) was expressed in all treated groups. However, MAP-2, as a mature neural marker, was expressed in CSR group.
The findings suggest that CSF accompanied RA lead to differentiation of cells with neuronal and glial phenotypes from BMSCs in vitro.