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目的:研究未经诱导SD大鼠骨髓间充质干细胞(MSCs)培养4周(4～6代)膜电流特性,为移植细胞的选取和细胞移植后心肌电生理研究奠定基础。方法:按文献方法获得和培养MSCs至第4周,利用全细胞膜片钳技术对培养4周的MSCs膜电流进行检测,以各种钾钠钙通道阻滞剂对电流成分进行分析鉴定。结果:本实验成功封接并有电流表达细胞共39例,所有检测到的电流均以相应阻滞剂证实。共有4种钾电流表达即缓慢激活延迟整流钾电流、瞬时外向钾电流、超速激活延迟整流钾电流和内向整流钾电流(Ikir),3种外向钾电流在+60mV时电流峰值均值分别为(0.3894±0.1230)nA、(0.4135±0.1029)nA、(0.2570±0.1135)nA;Ikir在-120mV时电流大小为(0.9613±0.1484)nA;此外在极少数MSCs上检测到河豚毒素敏感的内向钠电流(ITTX.Na),未检测到内向钙电流。结论:培养4周后的MSCs存在钾钠电流的复合表达,从电生理角度看,MSCs有分化成功能性心肌样细胞的离子基础,其可作为体内移植较理想细胞选择。
OBJECTIVE: To study the membrane current characteristics of cultured rat bone marrow mesenchymal stem cells (MSCs) for 4 weeks (4-6 generations) without inducing, and to establish the basis for the selection of transplanted cells and the electrophysiological study of myocardial cells after transplantation. METHODS: MSCs were harvested and cultured by literature method until the 4th week. Whole-cell patch-clamp technique was used to detect the membrane current of MSCs cultured for 4 weeks. The current components were identified by various potassium-sodium channel blockers. Results: The experiment successfully sealed and current-expressing cells in 39 cases, all detected currents were confirmed with the corresponding blockers. There were four kinds of potassium currents, namely, slow activation of delayed rectifier potassium current, transient outward potassium current, overstimulated delayed rectifier potassium current, and inward rectifier potassium current (Ikir). The peak currents of the three outward potassium currents at + 60mV were (0.3894 ± 0.1230) nA, (0.4135 ± 0.1029) nA, (0.2570 ± 0.1135) nA; Ikir was (0.9613 ± 0.1484) nA at -120mV; in addition, tetrodotoxin-sensitive inward sodium current ITTX.Na), no inward calcium current was detected. CONCLUSIONS: After 4 weeks of culture, MSCs present compound expression of potassium current, and from the electrophysiological point of view, MSCs differentiate into the ionic basis of functional cardiomyocyte-like cells, which can be used as ideal cell selection in vivo.