Co-culturing of neurosphere-derived oligodendrocyte like cells by motorneurons derived from mesenchymal stem cells for production of myelin

Background and Objective: One of the approaches for treatment of demylination diseases is cell therapy. Stem cells have been used as a source for generating oligodendrocyte. In a previous communication, the oligodendrocyte-like cells (OLCs) were generated from bone marrow stromal cells (BMSCs) using preinducers (dimethyl sulfoxide and retinoic acid) and induction protocol (PDGF, bFGF, heregulin and T3). In this investigation, we tried to increase the yield of oligodendrocyte-like cells (OLCs) by inducing the neural stem cells generated from neurospheres-derived BMSCs.

Materials and Methods: In this research study, BMSCs of adult female rats were expanded and then induced to form neurospheres (NS) using B27, the epidermal growth factor (EGF) and the basic fibroblast growth factor (bFGF), and subsequent isolation of neural stem cells (NSCs). This was followed by induction of the NSCs into OLCs with heregulin, PDGF-AA, bFGF and T3. BMSCs, NS, NSCs and OLCs were characterized using immunocytochemistry for fibronectin, CD44, CD90, CD45, Oct-4, O4, Oligo2, O1 and MBP markers. PDGF receptor α (PDGFR-α), Olig2 and MOG expression were evaluated using RT-PCR.

Results: The BMSCs expressed CD44, CD90, CD106 and Oct-4 the NSCs were immunoreactive to nestin and NF68, while the OLCs were immunoreactive to O4, Oligo2 and O1. In differentiating OLCs, Olig2 and PDGFR-α mRNA was detected using RT-PCR technique.

Conclusion: Functional OLCs were generated from BMSCs-derived NS with high yield and were able to produce myelin after co-culture by motor neurons.