Retinal Multipotent Stem-Cell Derived “MiEye” Spheroid 3D Culture Model for Preclinical Screening of Non-viral Gene Delivery Systems

Abstract

Non-viral retinal gene therapy is a promising therapeutic approach towards the management of retinal degenerative diseases especially glaucoma. Current methods of in vitro preclinical screening of candidate nanoparticle systems in monolayer cell cultures are not reliable in predicting in vivo performance. In this paper, we describe the development of a multipotent stem-cell derived three-dimensional “mini-retina” culture model (MiEye) that aims to simulate an in vivo clinical model for more reliable gene delivery system screening. Through the utilization of multiplex gene expression profiling, we have shown that retinal stem cells can be differentiated in 3D culture to generate retinal neurospheres comprising of multiple retinal cell types. The 3D cell culture model combined with confocal microscopy imaging and fluorescence profiling techniques is a powerful tool as a retinal gene and drug delivery screening model.

Evaluation of transfection with gemini nanoparticles for retinal neuroprotection in a dynamic microfluidic cell culture system

“…The 3D culture was one of the solutions, and it is worth mentioning that our group succeeded in developing such systems. 22,23 Although the cell conditions in 3D culture more closely resemble the cell conditions in vivo, drug delivery system interaction with cells can show many differences when comparing the static drug delivery in the 2D and 3D methods. Therefore, a microfluidic cell culture system, such as in this study, serves to simulate the dynamic exposure of the cells to the delivery system in vivo.…”

Materiomically Designed Polymeric Vehicles for Nucleic Acids: Quo Vadis?

“…104,105 While several groups have employed tumor spheroid models for screening polymeric 106−108 and peptide vehicles 109 for gene delivery, it is unclear whether spheroid models can be easily established for other cell types, particularly to model gene delivery to complex organ targets, such as the retina. 110 Fluorescence plate readers, which are typically used to assess transfection outcomes, process hundreds of samples in wellplates, 111 and relies on automation of plate switching and liquid handling. 112 Despite improvements in plate reader technology, particularly the incorporation of Raman and FRET capabilities, 113,114 they remain restricted to homogeneous measurements.…”

“…A common criticism of cellular microarrays, microfluidic devices, and well-platestechniques reliant on culturing cells on planar substratesis that they do not recapitulate in vivo conditions faithfully. Tissue spheroids and 3D cell culture scaffolds have been proposed to improve the physiological relevance of high-throughput screening experiments. , While several groups have employed tumor spheroid models for screening polymeric − and peptide vehicles for gene delivery, it is unclear whether spheroid models can be easily established for other cell types, particularly to model gene delivery to complex organ targets, such as the retina …”