Abstract
Bone marrow mesenchymal stem cells (MSCs) are used as a cell source for cardiomyoplasty; however, the cellular electrophysiological properties are not fully understood. The present study was to investigate the functional ionic channels in undifferentiated mouse bone marrow MSCs using whole cell patch-voltage clamp technique, RT-PCR, and Western immunoblotting analysis. We found that three types of ionic currents were present in mouse MSCs, including a Ca(2+)-activated K(+) current (I(KCa)), an inwardly rectifying K(+) current (I(Kir)), and a chloride current (I(Cl)). I(Kir) was inhibited by Ba(2+), and I(KCa) was activated by the Ca(2+) ionophore A-23187 and inhibited by the intermediate-conductance I(KCa) channel blocker clotrimazole. I(Cl) was activated by hyposmotic (0.8 T) conditions and inhibited by the chloride channel blockers DIDS and NPPB. The corresponding ion channel genes and proteins, KCa3.1 for I(KCa), Kir2.1 for I(Kir), and Clcn3 for I(Cl), were confirmed by RT-PCR and Western immunoblotting analysis in mouse MSCs. These results demonstrate that three types of functional ion channel currents (i.e., I(Kir), I(KCa), and I(Cl)) are present in mouse bone marrow MSCs.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / pharmacology
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Animals
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Barium Compounds / metabolism
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Blotting, Western
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Bone Marrow Cells / drug effects
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Bone Marrow Cells / metabolism*
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Calcimycin / pharmacology
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Cell Size
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Cells, Cultured
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Chloride Channels / drug effects
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Chloride Channels / genetics
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Chloride Channels / metabolism*
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Chlorides / metabolism*
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Clotrimazole / pharmacology
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Intermediate-Conductance Calcium-Activated Potassium Channels / drug effects
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Intermediate-Conductance Calcium-Activated Potassium Channels / genetics
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Intermediate-Conductance Calcium-Activated Potassium Channels / metabolism*
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Ionophores / pharmacology
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Membrane Potentials
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Mesenchymal Stem Cells / drug effects
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Mesenchymal Stem Cells / metabolism*
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Mice
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Mice, Inbred C57BL
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Nitrobenzoates / pharmacology
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Patch-Clamp Techniques
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Potassium / metabolism*
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Potassium Channel Blockers / pharmacology
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Potassium Channels, Inwardly Rectifying / drug effects
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Potassium Channels, Inwardly Rectifying / genetics
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Potassium Channels, Inwardly Rectifying / metabolism*
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RNA, Messenger / metabolism
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Reverse Transcriptase Polymerase Chain Reaction
Substances
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Barium Compounds
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Chloride Channels
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Chlorides
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ClC-3 channel
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Intermediate-Conductance Calcium-Activated Potassium Channels
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Ionophores
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Kcnn4 protein, mouse
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Kir2.1 channel
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Nitrobenzoates
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Potassium Channel Blockers
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Potassium Channels, Inwardly Rectifying
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RNA, Messenger
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barium chloride
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Calcimycin
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5-nitro-2-(3-phenylpropylamino)benzoic acid
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Clotrimazole
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4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid
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Potassium