Pre-clinical Model of Retinoblastoma Vitreous Seeds

Abstract

Purpose : The biggest challenge in treatment of retinoblastoma (Rb) is the presence of vitreous seeds. Vitreous seeding has a poorer prognosis for ocular salvage as reflected by multiple Rb classification systems. There is a lack of in vitro models of Rb vitreous seeds. Here, we adapt our in vitro 3D culture system to recreate Rb seeds in a vitreous-like microenvironment for more in-depth study.

Methods : Monocultures and co-cultures of CFSE-fluorescently labeled Y79 cells and human primary retinal microvascular endothelial cells (hRECs) were 3D cultured using magnetic levitation to re-create Rb seeds in the vitreous. We were able to visualize the growth of seeds over time in vitro by confocal microscopy.

Results : We generated in vitro Rb seeds similar to those found in Rb patients. The cultures were kept for over 2-weeks. Moreover, we were able to perform histological assessment to compare the 3D generated and ex vivo isolated seeds.

Conclusions : We have generated a pre-clinical model to investigate Rb seeds in vitro by using magnetic 3D cultures. Our novel methodology of fluorescently labeling the cells prior to magnetization allows isolation of both cell types to further characterize cells within the Rb microenvironment.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.