Copelas L, Briggs M, Grossman W, Morgan J P
Pflugers Arch. 1987 Mar;408(3):315-7. doi: 10.1007/BF02181475.
The purpose of this study was to develop a method for attachment of single isolated cardiac myocytes to a transducer for recording isometric tension development. Cardiac myocytes were isolated from the hearts of the toad, Bufo marinus or ferrets by enzymatic digestion with collagenase. The method that we used provided a 60-80% yield of Ca++-tolerant cells. A suspension of cells was placed into a superfusion chamber coated with bovine thrombin. Two glass microtools - each attached to a micromanipulator - were brought into proximity with the ends of a single myocyte; one of the microtools was attached to the element of a low-level force transducer. Human fibrinogen was loaded into a fine-tipped glass micropipette mounted on a micromanipulator. Small amounts of fibrinogen were pressure-ejected from the pipette at each junction between the microtool and the end of the myocyte. The fibrin that formed produced a stable attachment of the ends of the myocyte to the microtools. The myocyte could subsequently be stretched and a length-tension curve recorded. We have used this method to record concentration-dependent tension development in response to the Ca++-ionophore, A23187, and potassium depolarization. Our results indicate that fibrin glue may facilitate the study of the mechanical properties of isolated myocytes.
本研究的目的是开发一种将单个分离的心肌细胞附着于换能器以记录等长张力发展的方法。通过用胶原酶进行酶消化，从蟾蜍（Bufo marinus）或雪貂的心脏中分离出心肌细胞。我们使用的方法可获得60 - 80%的耐Ca++细胞产量。将细胞悬液置于涂有牛凝血酶的灌流室中。两个玻璃微工具——每个都连接到一个显微操作器上——靠近单个心肌细胞的两端；其中一个微工具连接到一个低水平力换能器的元件上。将人纤维蛋白原装入安装在显微操作器上的细尖玻璃微量移液器中。在微工具与心肌细胞末端的每个连接处，从移液器中压力喷射出少量纤维蛋白原。形成的纤维蛋白使心肌细胞末端与微工具稳定附着。随后可以拉伸心肌细胞并记录长度 - 张力曲线。我们已使用此方法记录了对Ca++离子载体A23187和钾去极化的浓度依赖性张力发展。我们的结果表明，纤维蛋白胶可能有助于对分离心肌细胞力学特性的研究。
