Abstract 19614: Formation of 3D Cardiac Tissues From Human Pluripotent Stem Cells Using a Highly Reproducible, One-Step Encapsulation Approach
Human induced pluripotent stem cells (hiPSCs) provide great potential for improving drug testing, facilitating myocardial repair, and investigating heart development. Currently, SC derived cardiomyocytes (CMs) are pre-differentiated from hiPSCs, dissociated, and combined with a biomaterial to form 3D cardiac tissues. This demands additional cell-handling, results in a high level of cell loss, and disrupts pre-existing cell-cell junctions. Here we describe a one-step approach to directly encapsulate and differentiate hiPSCs into 3D cardiac tissues. HiPSCs were combined with poly(ethylene glycol)-fibrinogen and photocrosslinked to form 200 μm thick tissues. Encapsulated hiPSCs (day -3) were cultured in their pluripotent state for three days where they proliferated. On day 0, cardiac differentiation was initiated. Tissues showed areas of contractions starting on day 7, resulting in synchronously contracting tissues which retained their function over three months. Sarcomeric α-actinin and gap junction protein connexin 43 (Cx43) were expressed by day 10; aligned sarcomeres (day 30) and mature gap junctions (Fig. 1a) developed over time. CM maturation increased significantly from day 10 to day 30, as quantified by the ratio of β over α myosin heavy chain gene expression; Cx43 expression increased from 0.5±0.3 (day 20) to 1.0±0.1 (day 30). Additionally, day 15 CMs showed 1:1 correspondence to electrical stimuli of up to 2 Hz. Finally, TEM of day 130 tissues showed structural features of mature CMs, including Z-bands, T-tubules, and intercalated discs (Fig. 1b). In summary, this one-step encapsulation approach successfully produces highly reproducible 3D cardiac tissues with mature and functional features. Furthermore, these physiologically relevant CMs can be used to produce printed microislands, macrotissues, spheroids, or single CMs, based on the desired downstream application.
Fig. 1. 3D encapsulation of hiPSCs produce functional cardiac tissues.
Author Disclosures: P. Kerscher: None. A.J. Hodge: None. J. Kim: None. I.C. Turnbull: None. D. Seliktar: None. C.J. Easley: None. K.D. Costa: None. E.A. Lipke: None.
This research has received full or partial funding support from the American Heart Association.
- © 2014 by American Heart Association, Inc.