TRPM8 channel role in mesenchymal stem cells osteogenic differentiation

Abstract

Mesenchymal stem cells (MSCs) have properties that make them a good model to use in regenerative medicine. MSCs proliferation and differentiation are complex processes involving the activation of several molecules that induces changes in intracellular calcium concentration. Different ion channels are responsible for calcium influx including TRP channels. The present study evaluates the expression and functional activity of TRPM8 channels in human mesenchymal stem cells. Cells were characterized by the expression of the specific surface markers CD90+, CD105+ and CD34− and by the ability to differentiate into adipogenic and osteogenic linage. Western blot, immunocytochemistry and flow cytometry were performed to determine TRPM8 protein expression. Patch clamp recordings and fluorescence imaging with Fluo‐4 AM were used to analyze channel activity. We found that the agonist menthol induces an increase in ionic currents which were incompletely abolished by treatment of the cells with the antagonist BCTC. Changes in intracellular calcium concentration were detected after addition of menthol, suggesting an effect induced by activation of TRPM8 channels. Alizarin red staining showed that the TRPM8 agonist Icilin increased osteogenic differentiation. On the contrary, the TRPM8 antagonist BCTC decreased cell differentiation. These results were confirmed by real time PCR with the marker Alkaline Phosphatase (ALP). Results from this study suggest that the activity of TRPM8 channels in mesenchymal stem cells is related with regulation of osteogenic differentiation.Support or Funding InformationThis work was supported by Departamento Administrativo de Ciencia, Tecnología e Innovación, COLCIENCIAS, Project: Evaluación del papel de canales TRPM8 en la diferenciación de células madre mesenquimales derivadas de tejido adiposo; Grant ID 120365843092.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.