Development of a drug screening platform based on engineered heart tissue.
BACKGROUND
Tissue engineering may provide advanced in vitro models for drug testing and, in combination with recent induced pluripotent stem cell technology, disease modeling, but available techniques are unsuitable for higher throughput.


OBJECTIVE
Here, we present a new miniaturized and automated method based on engineered heart tissue (EHT).


RESULTS
Neonatal rat heart cells are mixed with fibrinogen/Matrigel plus thrombin and pipetted into rectangular casting molds in which two flexible silicone posts are positioned from above. Contractile activity is monitored video-optically by a camera and evaluated by a custom-made software program. Fibrin-based mini-EHTs (FBMEs) (150 microL, 600 000 cells) were transferred from molds to a standard 24-well plate two hours after casting. Over time FBMEs condensed from a 12x3x3 mm gel to a muscle strip of 8 mm length and, depending on conditions, 0.2 to 1.3 mm diameter. After 8 to 10 days, FBMEs started to rhythmically deflect the posts. Post properties and the extent of post deflection allowed calculation of rate, force (0.1 to 0.3 mN), and kinetics which was validated in organ baths experiments. FBMEs exhibited a well-developed, longitudinally aligned actinin-positive cardiac muscle network and lectin-positive vascular structures interspersed homogeneously throughout the construct. Analysis of a large series of FBME (n=192) revealed high yield and reproducibility and stability for weeks. Chromanol, quinidine, and erythromycin exerted concentration-dependent increases in relaxation time, doxorubicin decreases in contractile force.


CONCLUSIONS
We developed a simple technique to construct large series of EHT and automatically evaluate contractile activity. The method shall be useful for drug screening and disease modeling.
Hansen
Arne
A
Department of Experimental and Clinical Pharmacology and Toxicology, Cardiovascular Research Center, University Medical Center Hamburg-Eppendorf, Germany.
Eder
Alexandra
A
Bönstrup
Marlene
M
Flato
Marianne
M
Mewe
Marco
M
Schaaf
Sebastian
S
Aksehirlioglu
Bülent
B
Schwoerer
Alexander P
AP
Schwörer
Alexander
A
Uebeler
June
J
Eschenhagen
Thomas
T
D000200:Action Potentials / Q000502:physiology; D000367:Age Factors; D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation* / Q000379:methods*; D008904:Miniaturization; D009206:Myocardium / Q000166:cytology*; D051381:Rats; D013678:Technology, Pharmaceutical / Q000295:instrumentation* / Q000379:methods*; D023822:Tissue Engineering / Q000295:instrumentation* / Q000379:methods*