Characterization of ion currents of murine CD117pos stem cells in vitro and their modulation under AT2R stimulation.

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Title: Characterization of ion currents of murine CD117pos stem cells in vitro and their modulation under AT2R stimulation.
Authors: Ludwig, M., Skorska, A., Tölk, A., Hopp, H.‐H., Patejdl, R., Li, J., Steinhoff, G., Noack, T.
Source: Acta Physiologica. Jul2013, Vol. 208 Issue 3, p274-287. 14p. 1 Color Photograph, 8 Graphs.
Subjects: Hematopoietic stem cells, C-kit protein, Cardiac regeneration, Angiotensin II, Angiotensin receptors, Reverse transcriptase polymerase chain reaction
Abstract: Aim Hematopoietic stem cells, especially CD117pos cells, have been found to possess a regenerative potential in various tissues, in particular cardiac muscle. However, the characterization of the relevant ion currents of stem cells prior to implantation lacks documentation. Activation of angiotensin II type 2 receptor ( AT2R) can lead to further cell differentiation and receptor auto-expression and might thus influence electrophysiological properties of CD117pos stem cells. This study was designed to functionally characterize membrane currents of CD117pos cells under normal and AT2R-stimulated conditions. Methods CD117pos murine bone marrow stem cells were isolated with MACS technique and stimulated for the AT2R with angiotensin II and losartan for 3-5 days prior to patch-clamp measurements. RT- PCR was used to determine channel expression. Endothelial properties were analysed with immunocytochemistry and ac LDL uptake assay. Results A well-expressed inward rectifying current ( IKir) was identified in cultured CD117pos cells. Furthermore, a ZD 7288 ( HCN channel blocker)-sensitive current component was isolated. Voltage-dependent potassium currents and chloride currents were less expressed. A small fraction of cells demonstrated voltage- and time-dependent inward currents. In AT2R-stimulated cells inward rectifying the hyperpolarization-induced inward currents were slightly attenuated on the translational level but showed increased mRNA expression. Cultured CD117pos cells express CD31 and VEGFR-2 and significantly increased the uptake of ac LDL. Conclusions CD117pos cells do not have properties of action potential-generating cells and moderately change their excitability during AT2R stimulation. Electrophysiological and molecular properties of control and AT2R-stimulated cells point to a differentiation to vascular endothelial cells. This could increase beneficial vascularization in injured tissues. [ABSTRACT FROM AUTHOR]
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Database: Psychology and Behavioral Sciences Collection
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Abstract:Aim Hematopoietic stem cells, especially CD117pos cells, have been found to possess a regenerative potential in various tissues, in particular cardiac muscle. However, the characterization of the relevant ion currents of stem cells prior to implantation lacks documentation. Activation of angiotensin II type 2 receptor ( AT2R) can lead to further cell differentiation and receptor auto-expression and might thus influence electrophysiological properties of CD117pos stem cells. This study was designed to functionally characterize membrane currents of CD117pos cells under normal and AT2R-stimulated conditions. Methods CD117pos murine bone marrow stem cells were isolated with MACS technique and stimulated for the AT2R with angiotensin II and losartan for 3-5 days prior to patch-clamp measurements. RT- PCR was used to determine channel expression. Endothelial properties were analysed with immunocytochemistry and ac LDL uptake assay. Results A well-expressed inward rectifying current ( IKir) was identified in cultured CD117pos cells. Furthermore, a ZD 7288 ( HCN channel blocker)-sensitive current component was isolated. Voltage-dependent potassium currents and chloride currents were less expressed. A small fraction of cells demonstrated voltage- and time-dependent inward currents. In AT2R-stimulated cells inward rectifying the hyperpolarization-induced inward currents were slightly attenuated on the translational level but showed increased mRNA expression. Cultured CD117pos cells express CD31 and VEGFR-2 and significantly increased the uptake of ac LDL. Conclusions CD117pos cells do not have properties of action potential-generating cells and moderately change their excitability during AT2R stimulation. Electrophysiological and molecular properties of control and AT2R-stimulated cells point to a differentiation to vascular endothelial cells. This could increase beneficial vascularization in injured tissues. [ABSTRACT FROM AUTHOR]
ISSN:17481708
DOI:10.1111/apha.12115