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目的探讨60极微电极阵芯片(MEA)技术在整体心脏、心肌组织片和培养心肌等电生理研究中的应用。方法①80只豚鼠,开胸取心脏并以2.5ml/min速度Langendorff灌流心脏20min,信号稳定后采样;②60只SD大鼠,开胸取心,剪取5mm×5mm大小的心脏组织片,置于MEA盘,台氏液间歇灌流5min,用5μV×1ms的刺激脉冲连续刺激,刺激间隔为1s,比较心房和心室组织片场电位(FPs)形态和传导时间;③20窝出生后3天以内的昆明小白鼠,消化分离心肌细胞,培养于MEA盘内,进行组织信号采样。结果①大鼠整体心脏灌流30～90min内能够记录到稳定的自主节律下的FPs信号,心室肌场电位时程(FPdur)210±78ms;心房肌FPdur164±58ms;房室传导时间320±150ms;并能记录到FPs激动顺序图。②心肌组织靶点取样,台氏液间歇灌流和电刺激下,组织的兴奋性可以稳定保持2h。心室肌组织片FPdur115.80±11.61ms,心房组织片FPdur83.71±6.48ms,刺激信号在心房组织片的传导时间66.46±6.73ms,心室组织片传导时间47.40±5.62ms;③小鼠乳鼠心脏原代培养心肌24h时后开始有散在、不同步的细胞克隆搏动和点状FPs信号,72h后细胞融合,开始记录到基本同步的群动和多位点FPs信号,培养心肌自发搏动频率150±100次/分。结论 MEA技术可以稳定记录小动物整体灌流心脏,心脏定点取样组织片,培养心肌细胞60极场电位和电传导特性。
Objective To investigate the application of 60 microelectrode array chip (MEA) technology in electrophysiological study of whole heart, myocardial tissue and cultured myocardium. Methods ①80 guinea pigs were sacrificed and the heart was opened by thoracotomy and the heart was perfused with Langendorff at a speed of 2.5ml / min for 20min. The signal was stable and then sampled. ②60 SD rats were thoracotomized and the cardiac tissue pieces of 5mm × 5mm were cut and placed in MEA and Taisu liquid were perfusion for 5min, stimulated by stimulation pulse of 5μV × 1ms for 1s, and compared the shape and conduction time of field potentials (FPs) in atrial and ventricular tissues. ③The Kunming small Rat, digestion of isolated cardiomyocytes, cultured in MEA dish, the tissue signal sampling. Results ① The FPs signal of FPdur was recorded within 30-90 min after cardioplegia in rats. FP dur was 210 ± 78 ms; atrial FPdur was 164 ± 58 ms; atrioventricular conduction time was 320 ± 150 ms; And can record FPs excited sequence diagram. ② myocardial tissue sampling target, Tyrodes solution intermittent perfusion and electrical stimulation, tissue excitability can be stable for 2h. Ventricular myocytes FPdur115.80 ± 11.61ms, atrial tissue FPdur83.71 ± 6.48ms, the stimulation signal in the atrial tissue conduction time 66.46 ± 6.73ms, ventricular tissue conduction time 47.40 ± 5.62ms; ③ mouse suckling mice Myocardial cardiomyocytes were initially cultured for 24 h, then scattered and asynchronous cell cloning pulsation and punctate FPs signals were observed. After 72 h, the cells fused and began to record basic synchronous group and multi-site FPs signals to train the frequency of spontaneous beats ± 100 beats / min. Conclusion The MEA technique can stably record the whole perfusion heart and cardiac fixed-point tissue slices of small animals to cultivate the 60-pole field potential and electrical conduction characteristics of cardiomyocytes.