Li Gui-Rong, Sun Haiying, Deng Xiuling, Lau Chu-Pak
Department of Medicine, Research Centre on Heart, Brain, Hormones and Healthy Aging, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.
Stem Cells. 2005 Mar;23(3):371-82. doi: 10.1634/stemcells.2004-0213.
This study characterized functional ion channels in cultured undifferentiated human mesenchymal stem cells (hMSCs) from bone marrow with whole-cell patch clamp and reverse transcription polymerase chain reaction (RT-PCR) techniques. Three types of outward currents were found in hMSCs, including a noise-like rapidly activating outward current inhibited by the large conductance Ca(2+)-activated K(+) channel (I(KCa)) blocker iberiotoxin, a transient outward K(+) current (I(to)) suppressed by 4-aminopyridine (4-AP), and a delayed rectifier K(+) current (IK(DR))-like ether-à-go-go (eag) K(+) channel. In addition, tetrodotoxin-sensitive sodium current (I(Na.TTX)) and nifedipine-sensitive L-type Ca(2+) current (I(Ca.L)) were also detected in 29% and 15% hMSCs, respectively. Moreover, RT-PCR revealed the molecular evidence of high levels of mRNA for the functional ionic currents, including human MaxiK for I(KCa), Kv4.2 and Kv1.4 for I(to), heag1 for IK(DR), hNE-Na for I(Na.TTX), and CACNAIC for I(Ca.L). These results demonstrate that multiple functional ion channel currents--that is, I(KCa), I(to), heag1, I(Na.TTX), and I(Ca.L)--are expressed in hMSCs from bone marrow.
本研究采用全细胞膜片钳和逆转录聚合酶链反应(RT-PCR)技术,对培养的来自骨髓的未分化人骨髓间充质干细胞(hMSCs)中的功能性离子通道进行了表征。在hMSCs中发现了三种外向电流,包括一种类似噪声的快速激活外向电流,被大电导Ca(2+)激活K(+)通道(I(KCa))阻断剂iberiotoxin抑制;一种瞬时外向K(+)电流(I(to)),被4-氨基吡啶(4-AP)抑制;以及一种延迟整流K(+)电流(IK(DR))样的醚-去-去(eag)K(+)通道。此外,分别在29%和15%的hMSCs中检测到了河豚毒素敏感的钠电流(I(Na.TTX))和硝苯地平敏感的L型Ca(2+)电流(I(Ca.L))。此外,RT-PCR揭示了功能性离子电流相关mRNA高水平表达的分子证据,包括I(KCa)的人MaxiK、I(to)的Kv4.2和Kv1.4、IK(DR)的heag1、I(Na.TTX)的hNE-Na以及I(Ca.L)的CACNAIC。这些结果表明,多种功能性离子通道电流,即I(KCa)、I(to)、heag1、I(Na.TTX)和I(Ca.L),在来自骨髓的hMSCs中表达。
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