Marotta Mario, Bragós Ramón, Gómez-Foix Anna M
Universitat de Barcelona, Barcelona, Spain.
Biotechniques. 2004 Jan;36(1):68-73. doi: 10.2144/04361ST01.
Excitability in muscle cells manifests itself as contractility and may be evoked by electrical stimulation. Here we describe an electrical stimulator device applicable to cells seeded on standard multiwell plates and demonstrate how it effectively stimulates synchronous contraction of skeletal muscle C2C12 cells without damaging them. The electrical stimulator of cultured cells (ESCC) consists of two connection cards and a network of platinum electrodes positioned in such way that each well in a row is uniformly stimulated. The ESCC may produce a range of outputs based on the stimulation parameters it receives from a commercial pulse generator and can be placed in a standard cell incubator, allowing for long-term stimulation as required for biochemical and molecular biological assays. We show that a 90-min stimulation of C2C12 myotubes at 50 V, 30 ms of pulse duration, and 3 Hz of frequency enhances glucose metabolism and glycogen mobilization while oppositely modulating the activity ratio of glycogen metabolizing enzymes. Thus, we demonstrate that long-term electrical stimulation of C2C12 myotubes with the ESCC results in contractility and metabolic changes, as seen in exercising muscle.
肌肉细胞的兴奋性表现为收缩性,并且可以通过电刺激诱发。在此,我们描述了一种适用于接种在标准多孔板上的细胞的电刺激装置,并展示了它如何有效地刺激骨骼肌C2C12细胞同步收缩而不损伤它们。培养细胞电刺激器(ESCC)由两张连接卡和一个铂电极网络组成,其放置方式使得一排中的每个孔都能受到均匀刺激。ESCC可以根据从商用脉冲发生器接收的刺激参数产生一系列输出,并且可以放置在标准细胞培养箱中,以便根据生化和分子生物学分析的需要进行长期刺激。我们表明,在50 V、30 ms脉冲持续时间和3 Hz频率下对C2C12肌管进行90分钟的刺激可增强葡萄糖代谢和糖原动员,同时反向调节糖原代谢酶的活性比。因此,我们证明,使用ESCC对C2C12肌管进行长期电刺激会导致收缩性和代谢变化,这与运动肌肉中的情况一致。