单个胃肠道平滑肌细胞的分离及其细胞膜离子通道检测

Dissociation of Single Gastrointestinal Tract Smooth Muscle Cell and Detection of Its Cellular Membrane Ion Channel

目的:建立胃肠道平滑肌细胞的分离及离子通道检测方法,为胃肠道动力性疾病和离子通道相关性疾病的研究提供技术平台。方法:酶消化法急性分离胃肠道平滑肌细胞,运用离子通道膜片钳技术在全细胞模式下检测单个平滑肌细胞膜上离子通道的状况。结果:用酶消化法急性分离可获得细胞膜完整生物活性状态良好的单个胃肠道平滑肌细胞,该细胞符合膜片钳实验的要求;运用离子通道膜片钳技术在全细胞模式下可以记录基础状态和干预后单个胃肠道平滑肌细胞细胞膜上离子通道的开放状况,可用于相关的病理、生理、药理研究。结论:酶消化急性分离法是获得单个胃肠道平滑肌细胞的一种简单、有效、易操作的方法,运用该方法结合离子通道膜片钳技术可以准确记录胃肠道平滑肌细胞细胞膜上离子通道的开放状况。

Objective： To establish a series of convenient methods for dissociating single gastrointestinal tract smooth muscle cell and detecting its cellular membrane ion channel. Methods： Single gastrointestinal tract smooth muscle cell was dissociated from muscle strips by using collagenase. Cellular membrane ion channel were detected by patch clamp technique under the whole-cell recording mode. Results： The single gastrointestinal tract smooth muscle cell which was dissociated by collagenase can maintain its physiological functions, and it can be used in patch clamp experiments. By using patch clamp technique, cellular membrane ion channel could be detected facilely. Conclusion： Dissociation by using collagenase is a convenient and reliable method to obtain single gastrointestinal tract smooth muscle cell, and patch clamp technique can be used to detect gastrointestinal tract smooth muscle cell cellular membrane ion channel punctually.