Engineered cardiac tissue microsphere production through direct differentiation of hydrogel-encapsulated human pluripotent stem cells.

Autor: Finklea FB; Department of Chemical Engineering, Auburn University, United States., Tian Y; Department of Chemical Engineering, Auburn University, United States., Kerscher P; Department of Chemical Engineering, Auburn University, United States., Seeto WJ; Department of Chemical Engineering, Auburn University, United States., Ellis ME; Department of Chemical Engineering, Auburn University, United States., Lipke EA; Department of Chemical Engineering, Auburn University, United States. Electronic address: elipke@auburn.edu.
Jazyk: angličtina
Zdroj: Biomaterials [Biomaterials] 2021 Jul; Vol. 274, pp. 120818. Date of Electronic Publication: 2021 Apr 20.
DOI: 10.1016/j.biomaterials.2021.120818
Abstrakt: Engineered cardiac tissues that can be directly produced from human induced pluripotent stem cells (hiPSCs) in scalable, suspension culture systems are needed to meet the demands of cardiac regenerative medicine. Here, we demonstrate successful production of functional cardiac tissue microspheres through direct differentiation of hydrogel encapsulated hiPSCs. To form the microspheres, hiPSCs were suspended within the photocrosslinkable biomaterial, PEG-fibrinogen (25 million cells/mL), and encapsulated at a rate of 420,000 cells/minute using a custom microfluidic system. Even at this high cell density and rapid production rate, high intra-batch and batch-to-batch reproducibility was achieved. Following microsphere formation, hiPSCs maintained high cell viability and continued to grow within and beyond the original PEG-fibrinogen matrix. These initially soft microspheres (<250 Pa) supported efficient cardiac differentiation; spontaneous contractions initiated by differentiation day 8, and the microspheres contained >75% cardiomyocytes (CMs). CMs responded appropriately to pharmacological stimuli and exhibited 1:1 capture up to 6.0 Hz when electrically paced. Over time, cells formed cell-cell junctions and aligned myofibril fibers; engineered cardiac microspheres were maintained in culture over 3 years. The capability to rapidly generate uniform cardiac microsphere tissues is critical for advancing downstream applications including biomanufacturing, multi-well plate drug screening, and injection-based regenerative therapies.
(Copyright © 2021 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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