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Justin B Lendermon, Nabil Saleh, Anderson Hudgens Webb, Bradley T Gao, Ryan P Lee, Matthew W Wilson, Jena J Steinle, Vanessa Marie Morales; Generation of in vitro intraocular tumor models for pre-clinical drug screening using nanotechnology. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):1660.
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© ARVO (1962-2015); The Authors (2016-present)
Three dimensional (3D) tumor cell cultures more closely resemble tumors in vivo, thus providing a better model for pre-clinical drug screening. This technology, Nano3D (Nano3D Biosciences, Inc., Houston, TX), was recently developed by Souza and colleagues to study glioblastomas. We adopted Souza’s techniques to generate 3D cell cultures of retinoblastoma and uveal melanoma for pre-clinical drug screening.
We used two retinoblastoma cell lines, Y79 and Weri, and three uveal melanoma cell lines, OMM1, 92.1 and Mel 270. First, we determined the minimum number of tumor cells to obtain significant data, maximizing their potential. Cells were co-cultured with nanoshuttle particles, made of iron oxide, to allow the formation of tumor spheroids upon contact with magnet in the dock station with capacity to record, at 37°C. Cells were dissociated and analyzed for cell viability and different surface markers. Then, we generated, in vitro, an extracellular matrix-like microenvironment to analyze tumor migration potential and disruption of extracellular matrix, in a similar fashion to the classical “scratch wound assay”. Third, we optimized tumor cell number and media volume to analyze cell growth by spheroid size.
Comparison of tumor growth for functional analyses in flat 2D surfaces versus the 3D nanotechnology system revealed significant differences. First, we found a more homogeneous phenotype in 2D flat surfaces than in the 3D. Second, we observed slight differences in adhesion molecules, specifically in UM, as they are adherent in 2D surfaces and not in the 3D.
The use of 3D cell culture is a powerful tool with potential applications for preclinical drug screening in intraocular tumors.
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