title: Generation of strip-format fibrin-based engineered heart tissue (EHT).
abstract: This protocol describes a method for casting fibrin-based engineered heart tissue (EHT) in standard 24-well culture dishes. In principle, a hydrogel tissue engineering method requires cardiomyocytes, a liquid matrix that forms a gel, a casting mold, and a device that keeps the developing tissue in place. This protocol refers to neonatal rat heart cells as the cell source; the matrix of choice is fibrin, and the tissues are generated in rectangular agarose-casting molds (12 × 3 × 3 mm) prepared in standard 24-well cell culture dishes, in which a pair of flexible silicone posts is suspended from above. A master mix of freshly isolated cells, medium, fibrinogen, and thrombin is pipetted into the casting mold and, over a period of 2 h, polymerizes and forms a fibrin cell block around two silicone posts. Silicone racks holding four pairs of silicone posts each are used to transfer the fresh fibrin cell blocks into new 24-well dishes with culture medium. Without further handling, the cells start to remodel the fibrin gel, form contacts with each other, elongate, and condense the gel to approximately ¼ of the initial volume. Spontaneous and rhythmic contractions start after 1 week. EHTs are viable and relatively stable for several weeks in this format and can be subjected to repeated measurements of contractile function and final morphological and molecular analyses.
authors: Department of Experimental Pharmacology and Toxicology, University Medical Center Hamburg Eppendorf (UKE), Hamburg, Germany.
authors: Sebastian
authors: S
authors: Schaaf
authors: Alexandra
authors: A
authors: Eder
authors: Ingra
authors: I
authors: Vollert
authors: Andrea
authors: A
authors: Stöhr
authors: Arne
authors: A
authors: Hansen
authors: Thomas
authors: T
authors: Eschenhagen
mesh_terms: D000818:Animals; D005337:Fibrin / Q000737:chemistry / Q000494:pharmacology*; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000187:drug effects*; D051381:Rats; D012685:Sepharose / Q000737:chemistry; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry
chemical_list: D005337:Fibrin; D012685:Sepharose