Protective effects of Human Bone Marrow-Derived Mesenchymal Stem Cells on Apoptosis Induced Human Neuroblastoma SH-SY5Y Cells

Abstract

Apoptotic cell death is important and anti-apoptotic strategies are crucial in treatment of neurological disorders including Alzheimer…s disease, Parkinson…s disease and stroke. Cell therapy using human bone marrow-derived mesenchymal stem cells (hMSCs) is an attractive tool to alleviate neurological disease We investigate whether culture media of hMSCs and coculted hMSC using by transwell have effects against apoptosis induced SH-SY5Ys. SH-ST5Ys were treated with 0.25？？M Stausrosporine(STS) for 24hrs to induce apoptosis. The media of hMSCs was collected after 2days of hMSC culture and added into STS-treated SH-SY5Ys. hMSCs plated in transwell were cocultured with STS-treated SH-SY5Ys. Cell viability was assessed by trypan blue dye exclusion method and MTS assay for 5days. Morphological, biochemical and Bcl-2 family western blot analysis were also performed. Number of viable cells was decreased progressively in STS-treated SH-SY5Ys in time, concentration-dependent manner and was not stopped with addition of new media including 10% FBS or media of hMSCs. Two days after application of transwell coculture system of hBM-MSCs, number of cells begin to increased significantly. Caspase-3 activity was maintained high and peaked at day 3 of addition of transwell hMSCs. Expression of bax was increased in STS-treated SH-SY5Y cocultured without hSMCs, but reversed progressively after hMSCs addition. Expression of Bcl-2 and surviving protein were increased in cocultured with hMSCs. These finding indicate that coculture with hMSCs protect apoptosis, but not only with addition of hMSCs culture media. Our results suggested that the hBM-MSCs might have an effect to induce intrinsic ‥self-repair… and regenerative capacity. These results might further support the potential therapeutic use of hBM-MSCs in neurological disease