Ps-120: Mouse Bone Marrow Mesenchymal Stem Cells (mBMSCs) Differentiation into Neural-like Cells by Staurosporine together with Morphine

Haramshahi SMA *, Azadbakht M , Kehtari M , Veisi-Raygani A ,


Objective: Mouse bone marrow mesenchymal stem cells (mBMSCs) can be differentiated into neural-like cells under appropriate experimental conditions. This study was designed to introduce a new protocol to neural differentiation based on staurosporine and morphine applications on neural differentiation in mBMSCs. Materials and Methods: Mouse BMSCs were isolated and cultured in DMEM containing 10% FBS. Differentiation assay and RT-PCR for rex-1 were done to affirmation multipotency of isolated cells. Cells were treated with 0, 50, 100, 214 and 316 nM of staurosporine (treatments I, II, III, IV and V; respectively) without morphine (group 1), and with 10-4 morphine (group 2). Neutral red uptake and total neurite length in each group were assessed 6, 12 and 24 hours after treatments. RTPCR for MAP-2 and immunostaining for GFAP, MAP-2 and Tub β-III were used to affirmation neural differentiation in BMSCs after treatment with differentiation medium containing staurosporine and morphine. Results: Cells were differentiated into osteocytes and adipocytes after exposure to special differentiation media. RT-PCR showed that isolated cells express rex-1 as a stem cell marker. Results showed that the viability of cells in group 2 in all treatments is higher than the same treatments in group 1 (p<0.05). Neurite outgrowth was increased in group 2 compared with group 1. In two groups viability of cells in all treatments was reduced from 6 to 24 hours but severity of viability decreasing in group 2 was lower than group 1(p<0.05). RT-PCR and immunostaining showed that the cells expressed neural markers after treatment with neural differentiation medium containing staurosporine and morphine. Conclusion: Our results suggest that staurosporine together with morphine can be used as a neural differentiation inducer in differentiation studies.