First Author: A.Laude SINGAPORE
Co Author(s): P. Shi Y. Tan W. Yeong
Back to previous
The retina is a complex multilayered tissue that can be affected by disease mechanisms such as age-related macular degeneration with associated retina pigment epithelium (RPE) and photoreceptor loss. We explore the use of three dimensional (3D) bioprinting to precisely deliver cells and biomolecules at fixed-point levels as a precursor to micro-tissues, micro-organs and memetic extracellular matrix models to investigate retinal diseases.
The photoreceptor/RPE bilayer was composed of cultured ARPE-19 (ATCC®CRL-2302™) and Y79 (ATCC®HTB-18™) that were put together using 3D bioprinting. The ARPE-19 cells were bioprinted as a monolayer onto a glass slide, followed by bioprinting of a second layer of Y79 cells. To compare, both cell types were also manually seeded separately on glass slides to act as controls. The bioprinted ARPE-19 and Y79 constructs were cultured for up to 14 days and investigated by reduction of presto blue, hematoxylin/eosin (HE) and immunohistological staining, confocal microscopy and scanning electrical microscopy (SEM).
The bioprinted ARPE-19 and Y79 cells showed robust biocompatibility. Moreover, the bioprinted ARPE-19 cells formed functional monolayer that demonstrated better cellular organization when compared to the control group. Further, Y79 cells can be precisely plotted at predesigned locations on the ARPE-19 cell monolayer to achieve controllable cell seeding density with assistance of the bioprinting technology. The ARPE-19 cells could form tight junctions upon formation of monolayer, while the Y79 cells robustly connected with the ARPE-19 monolayer when imaged using confocal microscopy, SEM and HE staining.
Bioprinting technology can be used to generate a retinal bilayer construct of ARPE-19 and Y79 cells which showed excellent cell viability and functionality up to 14 days of co-culture. This may potentially serve as a useful retinal model for studying retinal disease progression, drug mechanism and transplantation.