Various families of ion channels have been characterized in mesenchymal stem cells (MSCs), including some members of transient receptor potential (TRP) channels family. TRP channels are involved in critical cellular processes as differentiation and cell proliferation. Here, we analyzed the expression of TRPM8 channel in human bone marrow MSCs (hBM-MSCs), and its relation with osteogenic differentiation. Patch-clamp recordings showed that hBM-MSCs expressed outwardly rectifying currents which were increased by exposure to 500 μM menthol and were partially inhibited by 10 μM of BCTC, a TRPM8 channels antagonist. Additionally, we have found the expression of TRPM8 by RT-PCR and western blot. We also explored the TRPM8 localization in hBM-MSCs by immunofluorescence using confocal microscopy. Remarkably, hBM-MSCs treatment with 100 μM of menthol or 10 μM of icilin, TRPM8 agonists, increases osteogenic differentiation. Conversely, 20 μM of BCTC, induced a decrease of osteogenic differentiation. These results suggest that TRPM8 channels are functionally active in hBM-MSCs and have a role in cell differentiation.
<p>Este trabajo comporta un análisis crítico de la reparación del daño en el derecho administrativo colombiano. A partir del estudio de las nociones de ‘daño’ y ‘reparación’, esenciales para la atribución de responsabilidad civil, se demuestra el carácter limitado del entendimiento clásico de la responsabilidad extracontractual del Estado. Así pues, el artículo propone una comprensión de la reparación a partir del criterio de ‘universalización’, con independencia de las acciones por medio de las cuales se persiga una condena del Estado. Además, hace un estudio sobre los medios de control consagrados en el Código de Procedimiento Administrativo y de lo Contencioso Administrativo, y las acciones constitucionales a la luz de las diversas formas de reparación del daño.</p>
Due to their capacity to proliferate, migrate, and differentiate, mesenchymal stem cells (MSCs) are considered to be good candidates for regenerative medicine applications. The mechanisms underlying proliferation and differentiation of MSCs have been studied. However, much less is known about the mechanisms regulating the migration of MSCs. Platelet lysate (PL), a supplement used to promote cell expansion, has been shown to promote MSCs migration; however, the underlying mechanism are unknown. Here, by using adipose-derived rat MSCs (rMSCs) and the scratch assay in the absence and presence of various BK channels modulators, we evaluated the role of BK channels in mediating the PL-stimulated migration of rMSCs. We found that 5% PL increased rMSCs migration, and this effect was blocked by the addition of the BK channel selective antagonist Iberiotoxin (IBTX). In the absence of PL, the BK channel agonist NS1619, stimulated rMSCs migration to similar level as 5% PL. Addition of both NS1619 and 5% PL resulted in an increase in rMSCs migration, that was higher than when either one was added individually. From whole-cell recordings, it was found that the addition of 5% PL increased the magnitude of BK current density. By using Western blot and flow cytometry, it was found that PL did not affect the expression of BK channels. Together, our results indicate that as shown in other cell types, activation of BK channels by themselves also promote rMSC migration, and show that activation of BK channels contribute to the observed PL-induced increase in migration of rMSC.
El presente artículo muestra los resultados preliminares de un proceso de intervención cognitivo en el área de matemáticas para diferentes grupos de estudiantes que cursan los grados 7° a 9° en algunas Instituciones Educativas de los municipios de Pereira y Dosquebradas, actividad que atiende el primer objetivo formulado en el proyecto denominado “Didáctica para potenciar el pensamiento matemático mediante el uso de dispositivos lego” y que se refi ere a “Realizar un diagnóstico sobre el desarrollo del pensamiento matemático en estudiantes de la educación básica de las IE de los municipios de Dosquebradas y Pereira”.
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.
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