Multifunctional Cell-Culture Platform for Aligned Cell Sheet Monitoring, Transfer Printing, and Therapy

Abstract

While several functional platforms for cell culturing have been proposed for cell sheet engineering, a soft integrated system enabling in vitro physiological monitoring of aligned cells prior to their in vivo applications in tissue regeneration has not been reported. Here, we present a multifunctional, soft cell-culture platform equipped with ultrathin stretchable nanomembrane sensors and graphene-nanoribbon cell aligners, whose system modulus is matched with target tissues. This multifunctional platform is capable of aligning plated cells and in situ monitoring of cellular physiological characteristics during proliferation and differentiation. In addition, it is successfully applied as an in vitro muscle-on-a-chip testing platform. Finally, a simple but high-yield transfer printing mechanism is proposed to deliver cell sheets for scaffold-free, localized cell therapy in vivo. The muscle-mimicking stiffness of the platform allows the high-yield transfer printing of multiple cell sheets and results in successful therapies in diseased animal models. Expansion of current results to stem cells will provide unique opportunities for emerging classes of tissue engineering and cell therapy technologies.

Stretchable and Transparent Biointerface Using Cell‐Sheet–Graphene Hybrid for Electrophysiology and Therapy of Skeletal Muscle

“…Graphene layers on a copper foil are grown using chemical vapor deposition following protocols described in previous reports. [ 35,39 ] A 30 nm Ni layer is deposited onto the Si wafer and, subsequently, 7 nm/70 nm Cr/Au are deposited onto the Ni layer using thermal evaporation. Cr and Au layers are patterned by photolithography and wet etching.…”

“…To prepare cell-sheet-graphene hybrid, C2C12 myoblasts are seeded on the sample with a density of 5 × 10 4 cells cm −2 and cultured by using growth and differentiation media. [ 35 ] The formation of high quality biotic/abiotic interface enables the hybrid to electrically/optically stimulate and effectively monitor the electrophysiological characteristics in vitro and in vivo. Figure S2b, Supporting Information).…”

“…[ 35 ] The preferred adhesion of proteins and thereby cells to the graphene surface enhances efficiency of the cell culture. [ 35,41 ] The factor that quantifi es the softness of bioelectronics is the stretchability. Stretchable devices show superb deformability in bending, twisting, and stretching.…”

“…Serpentine-shaped, mesh-patterned graphene electrodes (≈50 µm width; Figure S1a, Supporting Information) are used to improve stretchability and softness of the system. The electrodes also monitor and promote proliferation and differentiation of muscle cells in vitro [ 35,36 ] as well as serve as an electrical stimulator and electrophysiology sensor in vivo. Au doping on the graphene mesh electrodes allows low impedance and high conductivity.…”

Current applications of graphene oxide in nanomedicine

“…However, ceria NPs are known to scavenge ROS and are therefore attractive candidates for inclusion into a graphene-based vehicles or TE platforms. 179 With this in mind, Kim et al 180 recently reported a graphene-based multifunctional platform that can suppress ROS generation. The multifunctional platform was capable of aligning plated cells and in situ monitoring of cellular physiological characteristics during proliferation and differentiation.…”