In vitro perfusion of engineered heart tissue through endothelialized channels
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In vitro perfusion of engineered heart tissue through endothelialized channels. / Vollert, Ingra; Seiffert, Moritz; Bachmair, Johanna; Sander, Merle; Eder, Alexandra; Conradi, Lenard; Vogelsang, Alexander; Schulze, Thomas; Uebeler, June; Holnthoner, Wolfgang; Redl, Heinz; Reichenspurner, Hermann; Hansen, Arne; Eschenhagen, Thomas.
in: TISSUE ENG PT A, Jahrgang 20, Nr. 3-4, 01.02.2014, S. 854-63.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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T1 - In vitro perfusion of engineered heart tissue through endothelialized channels
AU - Vollert, Ingra
AU - Seiffert, Moritz
AU - Bachmair, Johanna
AU - Sander, Merle
AU - Eder, Alexandra
AU - Conradi, Lenard
AU - Vogelsang, Alexander
AU - Schulze, Thomas
AU - Uebeler, June
AU - Holnthoner, Wolfgang
AU - Redl, Heinz
AU - Reichenspurner, Hermann
AU - Hansen, Arne
AU - Eschenhagen, Thomas
PY - 2014/2/1
Y1 - 2014/2/1
N2 - In engineered heart tissues (EHT), oxygen and nutrient supply via mere diffusion is a likely factor limiting the thickness of cardiac muscle strands. Here, we report on a novel method to in vitro perfuse EHT through tubular channels. Adapting our previously published protocols, we expanded a miniaturized fibrin-based EHT-format to a larger six-well format with six flexible silicone posts holding each EHT (15×25×3 mm³). Thin dry alginate fibers (17×0.04×0.04 mm) were embedded into the cell-fibrin-thrombin mix and, after fibrin polymerization, dissolved by incubation in alginate lyase or sodium citrate. Oxygen concentrations were measured with a microsensor in 14-day-old EHTs (37°C, 21% oxygen) and ranged between 9% at the edges and 2% in the center of the tissue. Perfusion rapidly increased it to 10%-12% in the immediate vicinity of the microchannel. Continuous perfusion (20 μL/h, for 3 weeks) of the tubular lumina (100-500 μm) via hollow posts of the silicone rack increased mean dystrophin-positive cardiomyocyte density (36%±6% vs. 10%±3% of total cell number) and cross sectional area (73±2 vs. 48±1 μm²) in the central part of the tissue compared to nonperfused EHTs. The channels were populated by endothelial cells present in the reconstitution cell mix. In conclusion, we developed a novel approach to generate small tubular structures suitable for perfusion of spontaneously contracting and force-generating EHTs and showed that prolonged perfusion improved cardiac tissue structure.
AB - In engineered heart tissues (EHT), oxygen and nutrient supply via mere diffusion is a likely factor limiting the thickness of cardiac muscle strands. Here, we report on a novel method to in vitro perfuse EHT through tubular channels. Adapting our previously published protocols, we expanded a miniaturized fibrin-based EHT-format to a larger six-well format with six flexible silicone posts holding each EHT (15×25×3 mm³). Thin dry alginate fibers (17×0.04×0.04 mm) were embedded into the cell-fibrin-thrombin mix and, after fibrin polymerization, dissolved by incubation in alginate lyase or sodium citrate. Oxygen concentrations were measured with a microsensor in 14-day-old EHTs (37°C, 21% oxygen) and ranged between 9% at the edges and 2% in the center of the tissue. Perfusion rapidly increased it to 10%-12% in the immediate vicinity of the microchannel. Continuous perfusion (20 μL/h, for 3 weeks) of the tubular lumina (100-500 μm) via hollow posts of the silicone rack increased mean dystrophin-positive cardiomyocyte density (36%±6% vs. 10%±3% of total cell number) and cross sectional area (73±2 vs. 48±1 μm²) in the central part of the tissue compared to nonperfused EHTs. The channels were populated by endothelial cells present in the reconstitution cell mix. In conclusion, we developed a novel approach to generate small tubular structures suitable for perfusion of spontaneously contracting and force-generating EHTs and showed that prolonged perfusion improved cardiac tissue structure.
U2 - 10.1089/ten.TEA.2013.0214
DO - 10.1089/ten.TEA.2013.0214
M3 - SCORING: Journal article
C2 - 24156346
VL - 20
SP - 854
EP - 863
JO - TISSUE ENG PT A
JF - TISSUE ENG PT A
SN - 1937-3341
IS - 3-4
ER -