Low frequency mechanical stimulation inhibits adipogenic differentiation of C3H10T1/2 mesenchymal stem cells

Oscillatory mechanical stimulation at relatively high frequencies ( 0.1Hz) has been shown to inhibit adipogenic and promote osteogenic differentiation of mesenchymal stem cells. However, for physiological interpretations and ease of implementation it is of interest to know whether different rates of mechanical stimulation can produce similar results. We hypothesized that relatively low frequency mechanical stimulation (0.01Hz) can in hibit adipogenic differentiation of C3H10T1/2 mouse mesenchymal stem cells, even in a potent adipogenic differentiation medium. C3H10T1/2 cells were cultured in adipogenic medium under control (non- mechanicallystimulated) conditions and under oscillatory surface stretch with 10% amplitude and 0.01Hz frequency for 6 h per day for upto 5 days. Cell population was assessed by counting and adipogenic differentiation was assessed by real- time quantitative PCR( qPCR) analysis of peroxisome proliferator- activated receptor gamma ( PPAR gamma) and fatty acid binding protein 4 (FABP4) after 3 and 5days. Involvement of the ERK signaling pathway was assessed by Western blot. Low frequency mechanical stimulation significantly decreased expression of PPAR gamma after 3 days and FABP4 after 3 and 5days versus non-stimulated culture. ERK signaling was decreased in mechanically-stimulated culture, indicating a role in the inhibition of adipogenic differentiation. Application of this study: Low frequency mechanical stimulation may provide a technically simple means for control of mesenchymal stem cell differentiation in cell-based therapies, particularly for inhibition of differentiation toward undesired adipogenic lineages. (C) 2012 International Society of Differentiation. Published by Elsevier B.V. All rights reserved.