A method for recording isometric tension development by isolated cardiac myocytes: transducer attachment with fibrin glue

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Abstract
The purpose of this study was to develop a method for attachment of single isolated cardiac myocytes to a transducer for recording isometric rension 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.