ABSTRACT
Human iPSC-derived cardiomyocytes offer infinite possibilities for cardiovascular drug discovery and regenerative
medicine, but only if they reflect the physiology of an adult human heart. Current protocols produce cardiomyocytes with
embryonic rather than adult profiles, severely limiting their utility for the adult population. Recent studies suggest that
bioengineering approaches aimed at re-creating environmental cues in a dish can lead to marked improvements in
cardiomyocytes’ maturation profiles. New technological tools are required to translate these findings into the practice.
To enable scientists in academia and biotech/pharmaceutical industry to generate more mature hiPSC-derived
cardiomyocytes on demand, we propose to design, engineer, fabricate, and validate a novel bioengineering hardware system
that will optically control the pacing of hiPSC-derived cardiomyocytes over extended periods of time to drive their
maturation toward the adult phenotype in an activity-dependent manner. The proposed maturation-promoting system will
utilize our proprietary graphene-mediated optical stimulation technology that allows fast, reversible, and physiologically
relevant activation of cells. Our technology has several critical advantages, as (1) it requires neither genetic modifications
(vs. optogenetics) nor wires/electrodes (vs. electric field stimulation), (2) it offers an uncharted flexibility for tunable light-
controlled cell stimulation protocols, and (3) it is compliant with high-throughput and scalability requirements.
The components of the proposed system (LightKick) will include graphene-coated microplates (G-plates), a multi-LED
unit to illuminate cells cultured in G-plates inside a cell incubator, and an external multiparametric programmable controller
to define illumination parameters. G-plates have already been developed and validated for optical stimulation and all-optical
assays. We will further optimize G-plates in terms of the substrate rigidity and micropatterning. The development of two
other components, together comprising an optical stimulation module, will represent the main engineering efforts of the
proposed project. To tailor this module for long-term optical stimulation, we will take into consideration recently developed
light emitters, optoelectronic properties of G-plates, biophysical properties of cardiomyocytes, and long-term cell culturing
conditions. Extensive technical capabilities of our optical stimulation module will offer (1) the scalability (by delivering
stimulation signals in few wells or few plates on demand); (2) wireless monitoring of the module and remote control of
optical pacing; and (3) the flexible tunability across the wide range of physiologically-relevant light illumination parameters
using a user-friendly interactive GUI software.
Importantly, long-term optical stimulation of hiPSC-derived cardiomyocytes enabled by the LightKick would be fully
compatible with many environmental cues that might play in the maturation process. The LightKick system will allow stem
cell experts (1) to develop efficient and individualized cell stimulation patterns for producing adult-like patient-specific
hiPSC-derived cells, and (2) to easily combine dynamic electric environmental cues with other environmental cues, further
improving multiparametric maturation-promoting protocols. We anticipate that the LightKick system will dramatically
simplify and, thus, accelerate the process of finding fundamental and practical answers to the maturation problem.
